AND 
VACCINE  THERAPY 

FOB.  VETERINARY  SURGEONS 


V.  M.  Scott 


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CLINICAL  BACTERIOLOGY  AND 

VACCINE  THERAPY  FOR  VETERINARY 

SURGEONS 


CLINICAL  BACTERIOLOGY 

AND 

VACCINE  THERAPY 

FOR    VETERINARY    SURGEONS 


WILLIAM  SCOTT,  F.R.C.V.S. 


LONDON 
BAILLIERE,    TINDALL    AND    COX 

8,  HENRIETTA  STREET,  COVENT  GARDEN 

1913 

{All  rights  reserved] 


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TO 

SIR   ALMROTH   E.   WRIGHT,  M.D.,  F.R.S. 

DIRECTOR   OF   THE    DEPARTMENT   OF   THERAPEUTIC   IMMUNIZATION, 
ST.    MARY'S   HOSPITAL,    LONDON 

AS   A    SLIGHT   TRIBUTE   TO    HIS    KMINENCE   AS    A 
VACCINE   THERAPEUTIST 


134543 


PREFACE 

Ir  Sir  Almroth  Wright's  prophecy — "the  physician  of 
the  future  will  be  an  immunizator  " — is  true  regarding  the 
future  attitude  of  the  practitioner  of  human  medicine 
towards  sero  -  vaccine  therapy,  I  believe  it  may  with 
equal  truth  be  applied  to  the  practitioner  of  veterinary 
medicine.  Certainly  there  is  as  much  scope  for  its 
application  in  the  latter  profession  as  in  the  former — nay, 
more  :  the  human  practitioner  has  difficulties  and  anxious 
problems  to  encounter  which  to  the  veterinary  practitioner 
are  unknown.  Every  medical  man  knows  with  what 
prejudice  his  patients  look  upon  any  new  line  of  treatment, 
and  particularly  so  if  such  treatment  is  being  driven  home 
at  the  point  of  the  hypodermic  needle.  For  the  sake  of 
his  own  reputation,  he  is  often  loath  to  rouse  those  latent 
feelings,  with  the  result  that  he  moves  very  cautiously — 
sometimes  too  cautiously — to  the  detriment  of  his  patient. 
Again,  a  doctor's  patients  are  all  possessed  of  a  highly 
developed  nervous  system,  where  the  influence  of  mind 
over  matter  is  very  pronounced,  and  where  emotions  and 
idiosyncrasies  are  greatly  in  evidence,  requiring  a  strong- 
personality  and  the  comforting  but  forcible  courage  of  the 
trusted  physician  to  quieten;  but  all  these  qualifications 
sometimes  avail  little,  the  patient  discarding  the  treatment 
without  fair  trial.  Or,  it  may  be  by  a  sheer  coincidence  that 
after  a  dose  of  vaccine  has  been  administered  complications 
in  the  course  of  the  disease  supervene,  and  although  they 
may  have  nothing  whatever  to  do  with  the  treatment, 
vaccine   therapy   is   blamed  and  condemned,   the   medical 


viii  PREFACE 

attendant  undergoes  a  harassing  time,  his  reputation  is 
at  stake,  and  he  may  perchance  lose  a  lucrative  client. 
The  veterinary  surgeon  is  in  a  much  happier  position,  he 
does  not  require  to  consult  his  patient,  study  his  whims, 
or  pander  to  his  capricious  fancies,  or  have  his  reputation 
held  up  to  ransom  if  all  does  not  go  quite  well  with  the 
feelings  of  his  patient  or  the  doubtful  desires  of  his  client. 
If  he  has  gained  the  confidence  of  his  client,  he  can  treat 
his  patient  in  whatever  manner  he  chooses;  all  that  is 
required  is  a  restoration  of  his  animal  to  good  health.  If, 
therefore,  vaccine  therapy  is  scientifically  sound — and  who 
will  deny  this  ? — if  its  results  warrant  its  application  in 
filling  a  gap  no  other  course  of  therapeutical  treatment 
can,  it  behoves  the  practitioner  in  his  clients'  interest  and 
his  patients'  welfare  to  take  up  the  treatment,  not  half- 
heartedly, which  will  never  lead  to  success,  but  giving  it 
wholly  and  fully  his  serious  consideration,  rigidly  taking 
note  of  every  detail,  and  benefiting  from  the  teachings 
of  each  individual  case.  In  the  whole  field  of  veterinary 
science  I  know  of  no  subject  which  possesses  such  oppor- 
tunities for  research  and  carries  with  it  such  active  and 
varied  interests  than  vaccine  therapy.  Every  practitioner 
can  be  an  immunizer,  and  to  show  him  that  he  can,  with 
a  little  diligent  effort  on  his  own  part,  was  the  primary 
object  I  had  in  view  in  writing  this  little  book.  I  am  fully 
alive  to  the  many  imperfections  it  contains,  and  I  know  in 
abler  hands  there  is  scope  for  a  much  better  work.  The 
great  portion  of  this  book  was  written  in  the  small  hours 
of  the  morning,  often  after  a  busy  and  exacting  day,  and 
to  those  who  know  the  responsibilities  and  disturbing 
influences  of  a  busy  practice,  which  are  by  no  means  con- 
ducive to  successful  literary  effort,  not  to  say  mental 
concentration,  I  am  sure  I  shall  not  appeal  for  their 
indulgence  in  vain.  I  have  endeavoured  to  introduce  as 
little  debatable  matter  as  possible  in  order  that  the  reader 
may  not  have  to  wade  thi^ough  unnecessary  detail. 

When  further  information  on  bacteriology  and  a  fuller 


PREFACE  ix 

description  of  the  symptoms  and  diagnoses  of  diseases  is 
required,  the  reader  must  be  referred  to  standard  works 
on  these  subjects,  the  primary  object  of  this  book  being 
the  making  and  the  administration  of  vaccines. 

An  exception,  however,  has  been  made  in  the  case  of 
two  diseases — namely,  tuberculosis  and  swine  fever — where 
a  fuller  descriptive  detail  has  been  gone  into,  owing  to 
the  prominence  given  to  these  two  affections  by  the  State 
and  the  veterinary  surgeon  of  the  present  day.  My 
grateful  thanks  are  due  to  Sir  Almroth  Wright  for  kindly 
admitting  me  to  the  Vaccine  Department  of  St.  Mary's 
Hospital,  where  I  obtained  much  useful  and  practical 
information ;  and  to  Dr.  Poels  for  so  courteously  giving  me 
access  to  the  State  laboratories  and  clearly  describing  the 
technique  of  sero-vaccine  therapy  as  practised  at  Rijks- 
seruminzichting,  Rotterdam.  I  am  also  indebted  to  Messrs. 
Rose  and  Carless,  Dr.  Hewlett,  Dr.  Emery,  and  Mr.  Jowett 
for  the  loan  of  the  various  photographs  appearing  through- 
out the  book.  For  the  illustrations  of  instruments  and 
apparatus  I  have  to  thank  Messrs.  Arnold  and  Sons  and 
Messrs.  Grallenkamp. 

Finally,  I  have  to  thank  Messrs.  Bailliere,  Tindall  and 
Cox,  for  their  courtesy  in  making  alterations  and  cheerfully 
carrying  out  suggestions  which  appeared  to  me  to  be 
necessary  as  the  work  was  going  through  the  press. 

W.  S. 
Friarn  House, 
Bridgewater, 

October,  1913. 


CONTENTS 


CHAPTER 
I. 
II. 
III. 

IV. 

V. 

VI. 

VII. 


VIII. 

IX. 

X. 

XI. 
XII. 


XIII. 

XIV. 

XV. 


XVI. 

XVII. 

XVIII. 


The  Laboratory  - 

Preparation  of  Culture  Media 

Cultivation  of  Bacteria 

Staining  Methods  and  Stains 

Identification  of  Bacteria — General  Principles   - 

Glass-AVork  Requisites  and  how  to  make  Them     - 

The    Protective    Elements   of   the   Blood,   which 

protect    the    Animal    Body    from    Pathogenic 

Bacteria 
Vaccines  and  their  Mode  of  Preparation  - 
The  Syringe      ...  . 

Phenomena    following    Active     Immunization     by 

Vaccines       ...... 

Serums  and  their  Mode  of  Preparation 

Special    Diseases,    caused    by    Specific    Bacteria, 

which   are   Suitable   for  Treatment   by   Sero- 

Vaccine  Therapy — Bacterial  Diseases  affecting 

the  Cutaneous  System       .... 
Bacterial  Diseases  affecting  Synovial  Joints 
Bacterial  Diseases  affecting  the  Abdominal  Organs 
Bacterial     Diseases    affecting    the    Circulatory 

System  .-._.. 

Bacterial  Diseases  affecting  the  Nervous  System 
Diseases  of  the  Respiratory  Organs 
Swine  Fever      ------ 


PAGE 

1 

15 

20 
25 

47 


91 

105 
114 

125 
2  36 
142 
186 


II. 
III. 
IV. 


APPENDICES 

Sero  -  Vaccine    Therapy    in    Holland    and    Other 
Countries     -  -  ... 

Additional  Notes         ... 
Weights  and  Measures  - 

Order  of  the  Board  of  Agriculture  and  Fisheries 

Bibliography     -  -  - 

Index       ------- 


194 

197 
201 
205 

217 
218 


LIST  OF  ILLUSTRATIONS 


PLATES 

rLATE  FACING   PAGE 

I.   Author's  Laboratory  Table  -  -         Frontispiece 

II.  A  Corner  in  the  Author's  Laboratory  -  -  -2 

III.  Normal  Cell  showing  Various  Side-Chains-             -             -  56 
First  Step  in  the  Formation  of  Antitoxin,  showing  Receptor 

and  Haptophore  United              -             -                          -  56 

IV.  Formation  of  many  New  Eeceptors,  linked  to  which  are  a 

Number  of  Haptophores             -             -             -             -  58 

Owing  to  the  Rapid  Increase  of  Receptors,  separation  of 
many  of  them  from  the  Cell  takes  place — Antitoxin 

begins  to  Form  -                          ...             -  58 

V.  Antitoxin  shown  Free  in  the  Blood              -             -             -  58 
Antitoxin  united  with  the  Haptophore,  thereby  protecting 

the  Cell  from  Toxic  Injury         -             -             -             -  58 

VI.  Standardization  of  Bacterial  Emulsion,  showing  Bacteria 

and  Red  Blood-Corpuscles  -  -  -  -74 

VII.  Staphylococci            ------  92 

Bacillus  coli  communis         -            -             -            -             -  92 

VIII.  Anthrax  Bacilli         -                          -                          -             -  134 

Bacillus  tetani           ._.---  134 

IX.  Micrococcus  catarrhalis— Human  Sputum  -             -             -  144 

Pneumococci             ------  144 

X.  Streptococcus  of  Strangles  -                          -  148 

Glanders  Bacillus     -                                                    -             -  148 

XL  Actinomycosis  Bovis             -----  166 

XII    Tubercle  Bacilli  in  Sputum              -                          -             -  176 


IN  THE  TEXT 


1.  The  Microscope 

2.  Incubator 

3.  Thermo  Regulator 


PAGE 

5 
8 
9 


xiv  LIST  OF  ILLUSTRATIONS 

FIG.  PAGE 

4.  Dry  Heat  Sterilizer      -                          -                          -  -  10 

5.  Moist  Heat  Sterilizer   -  11 

6.  Hand-Driven  Centrifuge           -  -  12 

7.  Water-Driven  Centrifuge          -  -  13 

8.  Potato  Finger  -                                                                 -  -  17 

9.  Plugged  and  Sloped  Medium  Tube      -                          -  -  IS 

10.  Platinum  Needles         -             -  -  20 

11.  Plugged  Stab  Medium  Tube     -  -  -21 

12.  Petri  Dish,  or  Plate      -                                       -  -  22 

13.  Friinkel's  Tube  for  Anaerobic  Cultivation        -             -  -  24 

14.  Wright  Spreader           -            ...             -             -  26 

15.  Wright  Spreader  mounted  on  Another  with  Sealing- Wax  -  26 

16.  Cornet  Forceps             -             -  27 

17.  Hanging-Drop  Preparation      -             -             -             -  35 

18.  Types  of  Growth  in  Stab  Culture         -  41 

19.  Types  of  Growth  in  Stab  Culture         -                          -  -  41 

20.  Surface  Elevation  of  Colonies  -             -             -             -  -  42 

21.  A  Simple  Capillary  Pipette      -             -                          -  -  48 

22.  Throttled  Capillary  Pipettes     -                                       -  -  49 

23.  Drawn-Out  and  Sealed  Test-Tube       -  '           -             -  -  50 

24.  Sealing  off  One  End  of  a  Glass  Tube  -  -  51 

25.  Wright's  Blood-Capsule            -             -             -            -  -  52 

26.  Wright's  Blood-Capsule            -             -             -             -  -  52 

27.  Wright's  Blood-Capsule            -             -             -             -  -  52 

28.  Slide  showing  Blood-Drop        -             -             -            -  -  53 

29.  Agglutination  of  Bacteria         -             -            -             -  -  59 

30.  Micro-organism,  Amboceptor,  and  Complement          -  -  62 

31.  Pipettes,    one    with    Rubber    Teat,    containing     Elements 

separated   by   Columns   of   Air,    and   thoroughly   mixed 

ready  for  incubating          -             -             -             -  -  65 

32.  Vaccine  Syringe            ...-.-  78 

33.  Serum  Syringe              -             -             -             -            -  -  78 

34.  Chart  showing  the  Effects  of  a  Suitable  Dose  of  Vaccine  -  81 

35.  Chart  showing  when  a  Dose  of  Vaccine  is  too  Small  -  82 

36.  Chart  showing  when  a  Dose  of  Vaccine  is  too  Large  -  -  82 

37.  Vessel  used  at  the  Serum  Institute  for  collecting  Blood  from 

the  Tail  of  an  Immune  Pig             ....  192 


CLINICAL    BACTERIOLOGY    AND 
VACCINE-THERAPY 


CHAPTER  I 

THE  LABORATORY 

This  should  be  a  fair-sized  room,  and  where  possible  only 
used  for  laboratory  work.  Ample  daylight  must  be 
admitted,  and  the  whole  ought  to  present  a  clean  appearance. 
The  walls  should  be  washed  with  a  light-coloured  hygienic 
wash  or  paint,  of  which  there  are  many. 

The  woodwork  should  be  painted  white,  as  also  the 
cabinets,  cupboards,  etc.,  and  the  floor  covered  with  a  plain 
linoleum. 

As  it  is  essential  for  media,  stains,  etc.,  and  even  the 
microscope  itself,  to  be  kept  at  a  fairly  even  temperature, 
the  laboratory  should  be  heated  in  the  winter  months  by 
hot-water  pipes  or  fires. 

The  bench  or  table  must  be  placed  in  the  brightest 
place  in  the  room,  for  preference  opposite  a  window. 

An  ordinary  table  will  answer  one's  purpose.  This 
should  have  a  blackened  top,  along  the  centre  of  which 
runs  a  piece  of  white  paper,  and  the  whole  top  should  be 
covered  with  a  sheet  of  glass. 

Plate-glass,  of  course,  is  best,  but  on  a  large  table  it 
is  somewhat  expensive ;  and  provided  the  top  is  perfectly 
level,  ordinary  sheet-glass  will  answer  our  purpose. 

The  back  of  the  table  ought  to  carry  a  rack  for  holding 

l 


•2  Or-J-NJCAL  BACTERIOLOGY  AND 

stains,  etc.,  and  be  within  easy  reach  of  the  operator. 
These  stains  should  be  in  glass  bottles,  with  rubber  teat 
droppers  fixed  in  them,  and  labelled. 

The  microscope  is  conveniently  placed  in  the  middle  of 
the  table,  and,  of  course,  accessible  to  the  operator,  or  in 
the  best  position  for  reflecting  the  light  from  the  window, 
a  hanging  gas-bracket,  or  an  electric  pendant. 

Other  accessories  found  upon  the  table  should  be  a  test- 
tube  rack,  cornet  forceps,  platinum  needles,  forceps,  watch- 
glass,  various  shaped  and  sized  pipettes,  spirit  lamp,  slide 
trough  for  staining,  distilled  water  flask  and  dropper,  a 
slab  of  plasticine,  and  one  or  two  glass  dishes  with  covers 
to  them. 

Another  important  addition  is  a  laboratory  sink.  Any 
ordinary  sink  will  answer  if  it  is  deep  enough,  and  hot  and 
cold  water  should  be  laid  on,  with  an  additional  small  tap 
under  which  is  fixed  a  bracket  shelf,  and  upon  which  the 
glass  slide  to  be  washed  is  placed  for  the  removal  of 
excess  of  stain,  etc. 

Two  or  three  rows  of  shelves  should  be  fixed  upon  the 
wall  to  carry  bottles,  etc. 

A  pair  of  balances  will  be  found  most  useful,  and  if  put 
in  a  cupboard  with  a  glass  door  and  fixed  upon  the  wall  at 
a  convenient  height  they  occupy  little  room,  and  are  within 
easy  access.  Another  strong  shelf  should  be  made  to  carry 
an  incubator  and  the  two  sterilizers  (i.e.,  moist  heat  and 
dry  heat).  Under  each  of  these  a  gas-tap  should  be  fixed, 
a  hole  bored  in  the  shelf,  through  which  a  rubber  tube 
passes,  and  leading  to  a  Bunsen  lamp,  which  is  used  for 
the  purpose  of  heating  these  apparatus. 

It  is  advisable  to  cover  the  surface  of  the  shelf  with 
sheet  metal  to  reduce  the  danger  of  fire. 

In  addition  one  must  have  glass  tubing,  one  or  two  glass 
funnels,  graduated  measures,  and  glass  flasks.  The  latter 
need  not  be  graduated,  as  they  are  somewhat  expensive. 
To  standardize  them,  fill  with  water  from  a  given  graduated 
measure,  and  with  a  grease  pencil  draw  a  ring  round  in 


VACC1NE-THEBAPY  3 

a  line  with  the  water-mark.  Various-sized  flasks  will  be 
found  useful,  ranging  from  100  c.c.  to  1,000  c.c.  When  the 
practitioner,  for  monetary  reasons,  is  compelled  to  limit 
his  expenditure  in  fitting  up  his  laboratory,  it  is  astonish- 
ing how  he  can  with  a  little  ingenuity  economize.  For 
example,  a  Thermos  flask  may  take  the  place  of  an  incu- 
bator, an  opsonizer,  and  moist-heat  sterilizer,  the  only 
extra  required  being  a  thermometer,  which  is  passed 
through  the  cork.  Then,  again,  a  cheap  dry-air  sterilizer 
can  be  improvised  by  using  an  ordinary  baking  oven,  and 
suitable  for  sterilizing  glass  ware,  instruments,  etc. 

The   following  is  a  list  of  requisites   necessary  for  the 
laboratory,  with  an  improvised  addition  : 


Improvised. 
Thermostat. 


Incubator  and  opsonizer.   ) 

Sterilizer  (moist),  Koch's.  J 

Sterilizer  (dry) .  Baking  oven. 

Centrifuge  (hand,  water, 
electric) . 

Pair  of  scales. 

Thermometers  (one register- 
ing up  to  100°  C,  and 
one  to  300°  C). 

Oil  sterilizer.  Soup  ladle  fixed  on  stand. 

Vaccine  syringe.  Hypodermic  syringe 

Bunsen  burner.  Blow  lamp. 

Spirit  lamp. 

Tubing      (glass     and 
rubber). 

Test-tubes. 

Slides. 

Cover-glasses. 

Flasks,  funnels. 

Watch-glasses. 

Cedar-oil. 

Canada  balsam. 


4  CLINICAL  BACTERIOLOGY 

Platinum  needles  (straight 

and  looped). 
Filter-paper. 
Glass-cutting  knife. 
Pipettes. 
Teats  (rubber). 
Plasticine. 

Emery-paper  (Hubert's  00) . 
Cotton-wool. 
Bottles,  stock  vaccines. 
Bottle  of  lysol. 
Etc. 

The  Microscope. 

For  bacteriological  work  a  sound  microscope  is  essential. 

It  must  have  a  firm,  rigid  stand,  and  an  accurate  fine  and 
coarse  adjustment. 

It  should  be  fitted,  or  be  capable  of  being  fitted,  with  sub- 
stage  condenser,  mechanical  stage,  objectives  f,  \,  and  -j^, 
oil  immersion,  and  two  eyepieces  giving  a  magnification 
up  to  1,000  diameters.  When  higher  magnifications  are 
required,  it  is  a  question  of  employing  extra  eyepieces 
which  possess  higher  magnifications  than  those  usually 
supplied  with  an  ordinary  bacteriological  microscope.  These 
can  be  obtained  from  any  of  the  leading  microscope-makers. 

A  comparatively  new  and  important  accessory  for  bac- 
teriological work  is  the  dark-ground  condenser  constructed 
for  observation  of  living  bacteria  in  an  unstained  condition. 
Under  this  mode  of  illumination  no  staining  is  required, 
and  the  bacteria  are  so  illuminated  that  they  appear  as 
self-luminous  bodies  upon  a  dark  background. 

The  microscope  we  possess  is  a  Leitz,  latest  model,  and 
is  fitted,  as  above  described,  with  an  additional  continuous 
safety  micrometer  fine  adjustment,  which  prevents  any 
chance  of  damaging  the  objectives  if  accidentally  over- 
focussed  and  brought  into  contact  with  the  specimen  slide, 
a  most  essential  point,  at  least  so  far  as  beginners  are  con- 


Fig.  1.— The  Microscope. 

This  model  does  not  illustrate  a  mechanical  stage,  which  is  a 
most  essential  accessory. 


6  CLINICAL  BACTERIOLOGY  AND 

cerned.  The  instrument  is  also  fitted  with  an  arched  limb, 
which  gives  a  large  working  space  above  the  object  stage, 
and  incidentally  acts  as  a  convenient  handle  for  lifting  the 
microscope  without  putting  any  extra  strain  upon  some  of 
the  other  vital  parts. 

The  stage  is  circular,  and  can  be  rotated  on  its  own 
axis;  it  is  also  fitted  with  a  centring  arrangement,  which 
is  an  advantage  when  making  an  examination  of  urinary 
deposits,  etc. 

The  mechanical  stage  is  detachable,  and  is  so  fitted  to 
the  limb  of  the  microscope  that  after  removal  it  can  be 
screwed  back  and  occupy  exactly  the  same  position  each 
time. 

The  instrument,  of  course,  inclines  to  a  horizontal  posi- 
tion, and  possesses  all  the  necessary  movements  for  ordi- 
nary bacteriological  purposes. 

The  Use  of  the  Microscope. 

Beginners  should  accustom  themselves  to  use  either  eye, 
and  the  unemployed  eye  should  remain  open  and  passive. 

Daylight  is  the  best  illuminant,  and  especially  so  if 
reflected  from  a  white  cloud.  Direct  sunlight,  of  course,  is 
useless.  As  the  majority  of  practitioners,  however,  of 
necessity  do  their  microscopical  work  when  the  light  of 
day  has  departed,  artificial  illumination  is  called  into  requi- 
sition. There  are  some  excellent  illuminants  made  by 
microscope-makers  for  this  purpose,  but  an  ordinary  gas  or 
oil  light  suffices  for  one's  needs. 

When  examining  an  object  with  the  dry  lens,  the  concave 
mirror  should  be  used,  which  converges  the  light  on  to  the 
object.  With  an  oil  immersion  lens  a  flat  mirror  should  be 
used ;  the  rays  reflected  are  converged  b}r  the  condenser 
and  brought  to  a  focus  on  the  object,  so  that  by  the  use  of 
the  condenser  a  considerable  amount  of  light  is  obtained. 
To  reflect  the  light  on  to  the  condenser,  the  mirror  should 
be  moved  about  until  a  suitable  angle  is  arrived  at. 

To  a  beginner  this  is  often  somewhat  difficult,  in  which 


VACCINE-THERAPY  7 

case  he  should  remove  the  eyepiece  and  look  down  the 
tube,  moving  the  mirror  until  he  sees  clearly  the  reflected 
light. 

We  will  now  presume  that  a  slide  has  to  be  examined 
under  a  TV  oil  immersion.  If  the  specimen  is  only  faintly 
stained,  it  is  well  to  make  a  ring  with  a  grease  pencil  round 
the  field.  This  serves  as  a  border  from  which  to  focus,  and 
is  most  useful.  A  drop  of  cedar-oil  should  be  placed  on 
the  specimen,  and  the  tube  lowered  by  the  coarse  adjust- 
ment until  the  objective  almost  touches  the  slide.  The 
beginner  should  now  place  his  eye  on  a  plane  with  the 
stage,  at  the  same  time  steadily  lowering  the  tube. 
Immediately  the  lens  touches  the  oil  it  will  appear  to  rise 
to  meet  the  objective.  With  the  fine  adjustment  then 
carefully  focus  until  the  objects  on  the  field  become  well 
defined. 

If  the  specimen  contains  bacteria  in  irregular  distribu- 
tion, it  is  advisable  to  make  one's  examination  by  way  of 
locating  the  best  area  with  a  low  power  first ;  in  fact, 
many  bacteriologists  carry  out  the  process  as  a  routine 
practice.  It  may  be  added  an  oil  immersion  lens  transmits 
two  or  three  times  more  light  than  a  dry  lens  of  the  same 
power,  while  the  Abbe  condenser  collects  the  rays  of  light 
reflected  from  the  mirror,  and  concentrates  them  at  a 
point  about  2  millimetres  above  its  upper  surface. 

If  the  nozzle  of  the  objective  requires  cleaning — and  this 
should  be  done  fairly  often — the  oil  can  be  removed  by 
xylol,  turpentine,  or  benzine  on  a  piece  of  chamois  leather. 

The  whole  instrument  should  be  kept  free  from  dust  and 
other  accumulations,  and  when  not  in  use  should  be  covered 
with  a  glass  shade. 

The  Incubator  and  Incubation. 

To  grow  bacteria  outside  the  animal  body  some  form  of 
incubator  is  required.  This  consists  of  a  metal  box  with 
double  walls,  the  space  between  which  is  filled  with  water, 
and  the  whole  encased  in  some  non-conductor,  such  as  felt 


8  CLINICAL  BACTERIOLOGY  AND 

or  wood.  The  water  is  heated  at  the  bottom  of  the  in- 
cubator by  a  gas,  oil,  or  electric  lamp.  The  top  of  the 
incubator  has  three  circular  holes  in  it,  one  for  the  insertion 


Fig.  2. — Incubator. 


of  a  thermo  regulator,  one  for  the  thermometer,  and  a  third 
for  the  filling  of  the  water  chamber.  The  thermo  regulator 
is  composed  of  a  mercurial  bulb  which  passes  through  a 


VACCINE-THERAPY 


9 


cork  at  the  top  of  the  incubator,  entering  the  air  chamber. 
The  mercury  rises  and  cuts  off,  or  rather  limits,  the  supply 
of  gas  entering  the  burner,  and  so  regulates  the  heat- 
supply.  The  regulator  in  the  writer's  possession  is  in  itself 
regulated  by  a  small  screw,  so  that  the  temperature  may 
be  set  at  any  desired  degree.  The  incubator  should 
be  provided  with  double  doors,  the  inner  one  of  which 
should  be  made  of  glass.  The  cultivation  of  bacteria 
usually  takes  place  in  the  laboratory  at  two  temperatures— 
i.e.,  20°  C.  (68°  F.),  suitable  for  those  germs  which  grow  out- 


Fig.  3. — Thermo  Regulator. 

side  the  animal  body;  and  37°  C.  (98-6°  F.),  or  body  tem- 
perature. All  media  are  suitable  for  the  higher  temperature 
save  gelatin,  which  melts  at  about  25°  C. 

It  must  also  be  remembered  that  bacteria  do  not  grow 
well  in  sunlight ;  they  should  therefore  be  incubated  in  the 
dark. 


Sterilizers  and  Sterilization. 

There  are  two  kinds  of  sterilizers  in  use — i.e.,  dry  and 
moist. 


10 


CLINICAL  BACTERIOLOGY  AND 


A"- dry-heat  sterilizer  consists  of  a  double-jacket  square  box, 
usually  made  of  strong  sheet  steel,  with  a  copper  bottom  and 
two  movable  perforated  shelves.  On  the  top  is  a  regulating 
slide  and  two  holes,  one  for  the  insertion  of  a  thermometer, 
and  the  other  for  a  thermo  regulator.  The  sterilizer  is 
fixed  on  a  raised  metal  stand  about  a  foot  in  height,  the 
heat  being  supplied  by  a  Bunsen  burner  or  by  a  power- 
ful oil  lamp.  The  articles  sterilized  by  dry  heat  are 
glass  flasks,  plates,  test-tubes,  pipettes,  cotton-wool,  etc. 


Fig.  4.— Dry  Heat  Sterilizer. 


To  completely  sterilize  any  of  these,  a  temperature  of 
150°  C.  must  be  maintained  for  three-quarters  of  an  hour. 
When  glassware  is  being  sterilized,  care  must  be  taken  to 
allow  the  sterilizer  to  cool  before  removing  any  of  its  con- 
tents; otherwise  they  are  liable  to  crack  when  exposed  to 
a  lower  outside  temperature. 

Steam  or  moist  -  air  sterilizers  consist  of  a  water-bath 
over  which  is  a  cylinder  with  a  perforated  diaphragm  fixed 


VACCINE-THERAPY 


11 


6  inches  from  the  bottom.  Several  varieties  are  in  use. 
The  one  we  use  is  Koch's  pattern,  and  answers  all  practical 
purposes.  A  steam  sterilizer  is  used  for  sterilizing  culture 
media. 

To  sterilize  media  thoroughly,  it  must  be  exposed  for  half 
an  hour  daily  for  three  successive  days.    The  first  steaming 


Fig.  5.— Moist  Heat  Sterilizer  (Koch). 

destroys  all  developed  bacteria,  the  second  destroys  the 
spore  after  it  has  developed  into  a  germ,  and  the  third  kills 
any  bacteria  which  may  not  have  developed  from  a  spore 
during  the  first  interval. 

The  Centrifuge. 

This  is  a  most  essential  laboratory  accessory,  and  there 
are  a  variety  on  the  market,  but  whether  they  are  driven 
by  hand,  water,  or  electricity,  the  principle  is  the  same — 


12 


CLINICAL  BACTERIOLOGY  AND 


namely,  to  obtain  so  many  revolutions  per  minute,  varying 
from  2,500  to  5,000.  It  is  devised  for  the  separation  of 
the  solid  and  liquid  elements,  blood,  pus,  milk,  urine,  and 
other  fluids,  for  examination  purposes. 


Fig.  6. — Hand-driven  Centrifuge. 


The  heematocrite  attachment  and  tube-holders  for  pur- 
poses of  combined  lightness  and  strength  are  made  of 
aluminium. 

A  hand  centrifuge  is  conveniently  fitted  to  the  table 
(which  must  be  firm  and  rigid)  by  means  of  a  double- 
tongued  clip. 

Where   one  can  obtain  a   sufficient  pressure,    a   water- 


VACCINE-THERAPY 


IS 


driven  centrifuge  is  a  thing  much  to  be  desired,  and  which 
is  in  most  common  use. 

Centrif legalizing  Blood. — The  quantity  of  blood  required 
is  placed  in  a  blood-tube,  and  before  coagulation  takes 
place  is  mixed  with  citrate  solution  and  centrifugalized. 
The  red  cells  are  thrown  down  first  by  reason  of  their 
specific  gravity,  appearing  at  the  bottom  as  a  red  layer ; 


I 


Fig.  7. — Water-driven  Centrifuge. 


next  the  white  corpuscles  as  a  grey  layer ;  and  on  the  top 
will  be  found  the  liquor  sanguinis  comparatively  clear. 
To  complete  this  process  one  has  to  use  the  centrifugal 
force  from  ten  to  fifteen  minutes. 

Centrifugalizing  Urine. — To  centrif ugalize  urine,  specially 
graduated  tubes,  tapering  to  one  end,  are  used,  the  speed 
required  being  about  2,500  revolutions  per  minute. 

Where  one  is  searching  for  bacteria  in  limited  numbers, 


14  CLINICAL  BACTERIOLOGY 

it  will  be  found  good  practice  to  centrifugalize  the  urine  in 
a  large  tube  first,  then  pipette  the  top  fluid,  leaving  about 
1  c.c. ;  this  or  less  may  be  placed  in  a  small  precipitating 
tube  and  centrifugalized.  By  this  process  a  greater  con- 
centration is  obtained. 

Centrifugalizing  Milk. — Milk  may  be  centrifugalized  for 
the  purpose  of  isolating  a  variety  of  bacteria,  and,  of  course, 
the  most  important  in  cow's  milk  is  the  bacillus  of  tuber- 
culosis so  far  as  veterinary  surgeons  are  concerned. 

Mix  20  c.c.  of  milk  with  1  c.c.  of  a  50  per  cent,  solu- 
tion of  potash,  heat  in  a  water-bath  until  the  solution  turns 
brown,  add  20  c.c.  of  acetic  acid.  Shake  the  whole  well, 
heat  in  water-bath  for  two  minutes,  and  centrifugalize  for 
ten  to  fifteen  minutes. 

The  top  fluid  is  now  poured  off ;  add  30  c.c.  of  hot  water 
to  the  sediment  and  centrifugalize ;  pour  off  the  super- 
natant fluid,  and  make  smears  on  three  slides ;  fix,  stain, 
and  examine. 


CHAPTER  II 

PREPARATION  OF  CULTURE  MEDIA 

At  the  present  time  a  large  variety  of  culture  media  are 
in  use,  but  for  general  routine  laboratory  work  the  list 
may  be  curtailed  to  five — i.e.,  (1)  broth;  (2)  agar-agar; 
(3)  gelatin;  (4)  blood-serum;  (5)  potato.  These,  except 
potato,  can  be  purchased  already  prepared  in  tabloid  form, 
but  for  economic  reasons  they  should  be  prepared  in  one's 
own  laboratory. 

Broth. 

This  is  easily  prepared,  and  it  forms  the  basis  for  the 
majority  of  other  media. 

Method. — Take  500  c.c.  of  water,  boil  well  for  half  an 
hour  in  a  double-contained  saucepan,  add  5  grammes  Lemco, 
10  grammes  peptone,  and  5  grammes  of  common  salt  ; 
then  boil  for  thirty  minutes.  Now  test  with  litmus-paper, 
and  it  will  in  all  probability  be  slightly  acid,  in  which  case 
a  solution  of  soda  should  be  added  drop  by  drop  until  the 
reaction  is  slightly  alkaline.  If  too  much  alkali  is  used, 
a  little  hydrochloric  acid  (diluted),  to  re-acidify,  should  be 
added,  and  then  again  neutralize.  Now  add  sufficient 
water  (some  having  been  lost  in  evaporation)  to  make  up 
1  litre,  and  filter  through  a  double  thickness  of  filter-paper 
into  a  500  c.c.  flask. 

Dry  the  mouth  of  the  flask  thoroughly  to  keep  the 
cotton-wool  from  sticking,  and  plug  firmly  with  absorbent 
wool.  Sterilize  at  60°  C.  for  half  an  hour  for  three  suc- 
cessive days. 


16  CLINICAL  BACTERIOLOGY  AND 

Gelatin. 

Method.— Take  100  c.c.  of  broth,  add  12£  grammes  of 
finely-cut  gelatin,  boil  thoroughly  until  all  is  dissolved, 
filter  into  a  flask  as  was  done  with  the  broth;  but,  as 
gelatin  solidifies  on  exposure  to  cold,  extra  heat  must  be 
applied  during  the  filtering  process.  The  flame  of  a  spirit 
lamp  placed  close  to  the  filter  funnel  answers  the  purpose. 
A  double-jacket  hot-water  funnel  can  be  purchased  to  serve 
the  same  object,  or  the  filter,  media,  and  flask,  may  be 
placed  in  an  oven  with  a  temperature  registered  at  40°  C. 

Gelatin  is  also  used  to  grow  bacteria  for  diagnostic  pur- 
poses, as  some  microbes  produce  a  digestive  ferment  and 
liquefy  gelatin,  and  others  do  not;  again,  some  liquefy 
rapidly,  and  others  slowly.  Gelatin  also  may  be  melted 
at  25°  C,  at  which  temperature  bacteria  are  not  destroyed. 
It  is  therefore  a  useful  medium  for  "  plating-out "  purposes. 

Agar-Agar. 

Method. — Take  100  c.c.  broth,  cut  up  2  grammes  of  agar 
into  very  fine  pieces,  soak  in  acetic  acid  (diluted). 

To  make  the  solution,  take  4  c.c.  glacial  acetic  acid,  add 
500  c.c.  water,  and  soak  for  fifteen  minutes ;  strain  off  the 
acid,  wash  the  agar  in  water  until  blue  litmus-paper  does 
not  become  red.  Now  place  the  broth  in  a  beaker  and  boil 
until  all  the  agar  has  been  dissolved.  Neutralize  with  an 
alkali,  and  allow  it  to  cool  to  about  50°  C.  To  make  the 
media  more  clear,  the  white  of  one  egg  to  each  500  c.c. 
of  media  should  be  added  before  boiling  again.  Filter  this 
gently  through  a  double  thickness  of  non-medicated 
surgical  lint  which  has  been  previously  moistened,  and 
prevent  the  media  from  solidifying,  as  was  done  in  the  case 
of  the  gelatin. 

Blood-Serum. 

This  medium  is  somewhat  difficult  to  prepare,  but,  fortu- 
nately, it  can  often  be  dispensed  with  altogether.  More- 
over, it  can  be  obtained  all  ready  for  laboratory  work. 


VACCINE-THERAPY  17 

Method. — Expose  the  carotid  of  a  horse  with  due  anti- 
septic precautions,  and  insert  a  sterile  cannula  into  the 
vessel.  The  blood  should  now  be  collected  in  a  sterile  jar 
by  means  of  a  rubber  tube  fixed  to  the  end  of  the  cannula, 
and  allowed  to  clot  for  twenty-four  hours. 

The  serum  is  then  drawn  off  with  sterile  pipettes,  placed 
in  sterile  test-tubes,  and  solidified  by  heating  to  70°  C, 
the  tubes,  of  course,  laid  on  their  sides  at  a  proper  angle  to 
give  the  necessary  slope. 

Potato. 

Method. — The  skin  should  be  well  scrubbed  and  peeled 
sufficiently  deep  to  remove  the  eyes;  cut  into  long  fingers, 
and  round  the  ends.  Now  divide  into  two,  cutting  obliquely, 
so  that  when  one  is  inserted  in  a  test-tube  it  will  have  the 


1 

Fig.  8.— (A)  Potato  Finger.     {B)  Cut  diagonally  ready  for 
Insertion  in  Tube. 


shape  of  a  sloped  gelatin  or  agar  medium  (Fig.  8).  Soak 
well  in  cold  water  for  twelve  hours,  put  each  into  a  test- 
tube,  plug,  and  sterilize  for  three-quarters  of  an  hour  on 
three  successive  days. 

Filling  and  Preparing  Test-Tubes  for  Cultivation. 

Having  prepared  the  medium,  we  require  sloped  and 
stabbed  test-tubes  ready  for  use.  Each  tube  should  be 
plugged  with  cotton-wool,  and  a  number  thus  prepared 
laid  out  on  the  table.  A  piece  of  glass  tubing  should 
be  procured,  on  one  end  of  which  a  rubber  pipe 
should  be  fixed,  the  glass  tubing  having  been  previously 
marked  to  allow  the  taking  up  of  a  given  quantity  of 
medium  each  time.     The  glass  end  should  now  be  inserted 


18 


CLINICAL  BACTERIOLOGY  AND 


into  the  hot  medium,  the  tubing  fixed  in  the  mouth  by  the 
teeth,  the  medium  sucked  up  to  the  volume  mark  and 
then  expelled  into  the  test-tube,  and  repeated  until 
all  the  tubes  have  been  charged.  Care  must  be  taken  not 
to  allow  any  of  the  medium  to  come  in  contact  with  the 
neck  of  the  tube,  in  which  case  the  cotton  plug  is  liable  to 
stick.  Another  method  is  to  procure  a  glass  funnel  to 
which  is  fixed  a  piece  of  rubber  tubing.     The  tubing  is 


4      Mi 


Fig. 


-Plugged  and  Sloped  Medium  Tube. 


clipped  by  a  spring  clip  (clasp  forceps  answer  the  purpose 
well),  and  the  whole  fixed  upon  an  iron  retort-stand.  The 
funnel  is  filled  with  the  medium,  and,  if  need  be,  kept  hot 
by  a  spirit  flame,  or  a  water-jacket  funnel  used.  The  tubing 
should  now  be  passed  down  the  test-tube  to  near  the 
bottom,  and  the  requisite  quantity  allowed  to  run  in  by 
loosening  the  grip  of  the  clip.     Here,  also,  care  has  to  be 


VACCINE-THERAPY  19 

taken  not  to  allow  the  medium  to  stick  on  the  mouth  or  neck 
of  the  test-tube. 

The  tubes  are  usually  charged  with  H  to  2  inches  of 
medium.  They  should  now  be  sterilized  in  a  Koch  sterilizer 
for  half  an  hour  on  three  successive  days,  and  laid  on 
their  sides  to  cool  to  give  a  sloped  surface,  or  upright  for  a 
stab  culture,  as  the  case  may  be. 


CHAPTER  III 
CULTIVATION  OF  BACTERIA 

Inoculating  Medium  Tubes. 

Having  chosen  our  culture  medium  upon  which  we  desire 
to  obtain  a  bacterial  growth,  either  with  the  idea  of 
assisting  or  confirming  our  microscopical  diagnosis  or  of 
obtaining  a  bacterial  emulsion  with  the  view  of  making  a 
vaccine,  we  sow  our  germ-laden  material  in  the  following 
manner : 

Take  a  platinum  loop  (this  consists  of  a  piece  of  glass 
tubing,  the  end  of  which  has  been  previously  softened  by 


3- 


Fig.  10. — Platinum  Needles. 

heat,  and  while  in  the  flame  a  piece  of  platinum  wire  is 
forced  into  the  soft  glass) ;  pass  it  through  the  flame  of  a 
spirit  lamp  to  sterilize  it.  Hold  in  the  left  hand  between 
first  and  second  fingers  a  medium  tube,  remove  the  cotton 
plug  and  place  it  between  the  ring  and  little  fingers  of  the 
same  hand,  taking  care  at  the  same  time  to  hold  the  tube 
(if  solid  medium)  mouth  downwards  to  prevent  ingress  of 
dust,  germs,  etc.  Dip  the  needle  into  the  morbid  material 
and  take  up  a  small  loopful;  pass  it  carefully  down 
the  tube  until  it  reaches  the  bottom,  letting  it  rest  gently 
20 


VACCINE-THERAPY 


21 


on  the  surface  of  the  medium,  and  draw  it  slowly  along 
the  whole  length,  care  being  taken  not  to  graze  the  surface 
with  the  loop  end.  Withdraw  the  needle  and  sterilize  it  as 
before.  Now  take  up  the  cotton-wool  with  forceps  and  set 
it  on  fire,  and  whilst  aflame  thrust  it  into  the  mouth  of 
the  tube ;  label  the  tube  with  grease  pencil — owner's 
name,  animal,  and  date — and  put  into  the  incubator. 

Where  we  desire  a  stab  culture,  the  process  adopted  is 


Fig.  11. — Plugged  Stab  Medium  Tube. 

the  same ;  but  in  this  case  we  pass  the  needle,  not  looped, 
but  pointed,  laden  with  the  pus  or  other  material,  through 
the  axis  of  the  medium,  give  it  a  slight  rotation,  withdraw, 
and  plug  as  before. 

Plate  Cultivation. 

Glass  plates  were  originally  used  for  cultivation,  but 
shallow  dishes  2  or  3  inches  in  diameter,  with  lids,  and 
known  as  Petri  dishes,  are  now  in  use. 


22  CLINICAL  BACTERIOLOGY  AND 

When  we  desire  to  obtain  pure  cultures,  bacteria  grown 
upon  test-tube  media  offer  great  difficulties.  To  overcome 
these,  plate  cultivation  is  adopted.  This  consists  of  several 
shallow  glass  plates,  each  provided  with  a  lid.  The  principle 
adopted  is  as  follows  :  When  suitable,  gelatin  is  usually 
employed,  but  agar  also  may  be  used.  Take  three  tubes 
of  gelatin  medium  and  melt  in  a  hot-water  bath.  Gelatin 
cools  to  25°  C,  and  agar  to  45°  C.  Mark  the  tubes  1, 
2,  3.  No.  1  is  inoculated  with  a  trace  of  the  material 
from  which  it  is  desired  to  make  the  pure  culture,  and 
completely  mixed  by  rolling  the  tube  between  the  hands,  so 
that  it  may  permeate  thoroughly  the  melted  gelatin.  Should 
this  mixture  be  plated  out  now,  however,  it  will  be  found 
to  be  too  rich  in  bacteria,  necessitating  their  adhering 
together  when  grown,  and  obscuring  their  characteristics. 


Fig.  12. — Petri  Dish,  or  Plate. 

To  obviate  this,  a  loopful  of  No.  1  tube  is  then  taken 
to  inoculate  tube  No.  2.  A  further  dilution  should  be 
made  by  inoculating  in  a  similar  manner  No.  3  from  No.  2, 
taking  up,  say,  a  half-dozen  loopfuls.  The  Petri  dishes 
having  been  previously  sterilized  in  a  dry-air  sterilizer, 
they  are  ready  for  use.  The  cotton-wool  is  now  removed 
from  the  tube,  and  the  mouth  sterilized  by  passing 
through  the  flame  of  the  spirit  lamp,  and  the  contents  of 
each  poured  into  separate  culture  plates,  the  lid  being 
slightly  raised  for  the  purpose,  and  put  aside  to  incubate. 
They  should  be  examined  daily  with  a  hand  lens  or  a  low 
power  of  the  microscope.  The  latter  can  be  done  by  tilting 
the  plate  and  placing  it  on  the  stage  of  the  microscope. 


VACCINE-THERAPY  23 

The  culture  in  plate  No.  1  will  usually  be  too  dense  to 
differentiate,  but  plates  Nos.  2  and  3  can  as  a  rule  be  used 
for  subculture.  This  is  done  by  taking  off  a  few  colonies 
with  the  platinum  loop  and  inoculating  a  test-tube  medium 
and  incubating  them.  Agar  plates  may  be  used  in  a 
similar  manner,  but,  owing  to  their  rapidly  -  cooling 
qualities,  the  work  must  be  done  very  quickly.  To  grow 
bacteria  with  the  view  of  procuring  a  large  supply  of 
bacterial  emulsion,  a  convenient  method  is  to  pour  the 
medium  into  flat-sided  bottles,  allow  it  to  set,  and  then 
pour  over  the  surface  a  small  quantity  of  bacteria-laden 
broth,  taking  care  it  is  evenly  diffused  all  over  the  face  of 
the  medium,  and  that  no  surplus  overflow  exists. 

Anaerobic  Cultivation. 

Many  bacteria  will  not  grow  in  the  presence  of  oxygen 
(free),  and  various  methods  are  adopted  to  remove  oxygen 
from  the  atmosphere  they  grow  in.  Some  of  these 
methods  are  very  simple,  others  very  complex.  The  simplest 
are  as  follows : 

1.  Take  a  narrow  test-tube  three-quarters  filled  with 
sterile  agar  medium  to  which  has  been  added  a  10  per 
cent,  solution  of  glucose,  place  it  in  hot  water,  and  boil 
well  for  five  or  ten  minutes.  Cool  and  make  solid  by  dip- 
ping in  cold  water.  The  germ-laden  needle  (a  large  one) 
should  now  be  thrust  into  the  axis  of  the  medium,  reaching 
the  bottom,  rotated  and  withdrawn,  and  the  sterile  plug 
inserted.  The  tube  is  then  heated  at  the  upper  border  of 
the  medium  to  seal  the  puncture,  and  a  well-fitting  rubber 
cap  applied.     The  tube  is  now  ready  for  incubating. 

2.  Another  simple  method  is  to  insert  a  plug  of  cotton- 
wool into  a  test-tube,  place  some  pyrogallic  acid  on  it  and 
moisten  with  caustic  soda  or  potash,  then  a  little  layer  of 
cotton-wool,  and  fix  on  the  mouth  of  the  tube  a  tight-fitting 
rubber  cap.  As  the  oxygen  becomes  absorbed,  the  cap 
will  be  drawn  in  by  suction. 

Four   grammes  of   pyrogallic  acid    and   8   grammes   of 


24 


CLINICAL  BACTERIOLOGY 


caustic  soda  dissolved  in  15  c.c.  of  water  will  absorb 
1,150  c.c.  of  oxygen. 

3.  The  following  is  more  complicated,  as  we  require 
coal  or  hydrogen  gas  to  replace  the  oxygen.  It  is  known 
as  "  Frankel's  method." 

A  stout  test-tube  has  inserted  into  its  mouth  a  rubber 
cork  having  two  perforations,  through  which  two  pieces  of 
glass  tubing  are  passed.    One  tube  reaches  into  the  medium  j 


Fig.  13. — Frankel's  Tube  for  Anaerobic  Cultivation. 


the  other  is  shorter  and  does  not.  Outside  the  test-tube  the 
tubes  are  bent  at  opposite  angles  to  each  other,  and  gradu- 
ally taper  to  points.  The  long  tube  is  connected  with  the 
hydrogen  supply,  and  the  small  tube  permits  of  its  exit. 
The  stream  of  gas  should  be  kept  up  for  several  minutes  at 
least,  to  permeate  the  whole  medium.  The  short  tube 
should  now  be  sealed  off  by  the  blowpipe  flame,  the  long 
tube  in  the  same  way,  and  the  joint  at  the  cork  covered 
with  paraffin. 


CHAPTER  IV 

STAINING  METHODS  AND  STAINS 

When  it  is  desired  to  make  a  microscopical  examination 
of  a  culture,  the  following  procedure  is  adopted  :  Place  on 
a  clean  slide  a  drop  of  distilled  water;  take  the  culture- 
tube  between  the  index  and  middle  finger  of  the  left  hand ; 
remove  the  cotton  plug  and  direct  the  mouth  of  the  tube 
downwards  if  the  media  is  solid ;  sterilize  a  platinum 
needle,  and  when  cool  pass  it  into  the  tube  and  pick  off  a 
portion  of  a  colony  with  the  loop,  taking  care  not  to  take  too 
much.  Now  heat  the  mouth  of  the  tube  with  the  flame ; 
scorch  the  cotton  plug,  and  thrust  it  into  the  test-tube.  The 
platinum  loop,  laden  with  bacteria,  should  be  well  mixed 
with  the  drop  of  sterile  water  previously  placed  at  one  end 
of  the  slide,  and  the  bacterial  emulsion  thus  made  spread  out 
with  the  platinum  needle  laid  flat  on  the  slide,  or  a  Wright 
spreader  may  be  used.    Such  a  spreader  is  made  as  follows : 

Wright's  Spreader. 

Take  the  thinnest  possible  slide,  make  a  nick  with  a 
glass-cutting  knife  *  about  halfway  along  its  side;  then  grasp 
the  two  ends  between  the  finger  and  thumb  of  either  hand, 
and,  advancing  the  thumb  of  the  right  hand  as  far  as,  or  a 
very  little  beyond,  the  intended  line  of  fracture  to  serve  as 
a  fulcrum,  break  the  slide  across  by  putting  a  transverse 
strain  upon  it,  at  the  same  time  exerting  a  pull  in  the 
longitudinal  direction.  An  arch  is  now  formed ;  the  point 
which  is  supported  by  the  thumb  will  correspond  to  the 

*  To  make  a  glass-cutting  knife,  see  p.  47. 
25 


26 


CLINICAL  BACTERIOLOGY  AND 


crown  of  the  arch.*  Cut  off  the  corners  by  nicking 
them  with  a  glass-knife,  and  mount  the  spreader  on  the 
back  of  another  slide,  fixing  it  with  sealing-wax. 


Fig.  14. — Weight  Spreader. 

The  concavity  of  the  spreader  so  made  will  allow  for  the 
passage  through  of  the  smaller  elements,  and  it  follows 
that  as  we  increase  the  angle  which  the  spreader  makes 


Fig.  15. — Wright  Spreader  mounted  on  Another  with 
Sealing  Wax. 

Note.— The  curve  has  been  exaggerated  to  make  the  description  more  clear. 

with  the  slide  we  proportionately  raise  the  height  of  the 
arch,  and  so  allow  the  larger  elements  to  pass  through. 

*  Sir  A  Wright's  "  technique  of  the  teat  and  capillary  glass  tube." 


VACCINE-THERAPY  27 

This  form  of  spreader  will  be  found  to  be  most  useful  in  the 
making  of  blood-films  and  in  counting  bacteria  against  the 
blood-corpuscles  when  we  desire  to  make  a  standardized 
emulsion.  When  dealing  with  pus  only,  it  is  necessary  to 
put  a  drop  on  the  end  of  a  slide  and  spread  it.  If  it  is  too 
thick,  a  little  distilled  water  or  normal  saline  solution 
should  be  added.  Needless  to  add,  to  get  a  perfectly  even 
spread,  a  clean,  grease-free  slide  should  be  used.  The  film 
so  made  is  now  allowed  to  dry,  and  fixed  by  heat  sufficient 
to  coagulate  the  albumin  and  to  retain  the  bacteria.  This 
heating  should  be  done  by  passing  the  slide  through  the 
flame  of  the  spirit  lamp  (specimen  face  upwards)  two  or 
three  times,  and  made  just  hot  enough  to  bear  comfortably 
on  the  back  of  one's  hand.  The  chosen  stain  is  now 
filtered  on  to  the  slide  through  filter-paper,  then  washed 


Fig.  16.— Cornet  Forceps. 

in  tap-water,  dried  with  filter-paper,  and  completely  dried 
over  the  flame.  In  the  same  way  cover-glasses  are  stained, 
holding  them  with  cornet  forceps. 

Usual  Method  of  Fixing  Films. 

1.  The  simplest  method  of  fixing  a  film  to  a  slide  or 
cover-glass  is  to  pass  it  through  the  flame  of  a  Bunsen 
burner  (specimen  side  uppermost)  three  or  four  times.  In 
this  case  the  albuminous  elements  are  coagulated  by 
heat. 

2.  Absolute  alcohol  fixes  a  specimen  in  five  to  ten 
minutes. 

3.  Corrosive  sublimate  (a  saturated  aqueous  solution) 
fixes  in  two  to  three  minutes,  and  even  less  in  some 
cases. 


28  CLINICAL  BACTERIOLOGY  AND 

4.  Formalin  (a  10  per  cent,  aqueous  solution)  fixes  in 
two  or  three  minutes. 

As  we  shall  see  (page  33),  where  Jenner's  or  Leishman's 
stains  are  used  no  fixing  is  required,  as  these  stains  are 
already  supplied  with  fixing  agents. 

Stains. 

Where  one  is  examining  pathological  fluids,  blood,  etc., 
for  bacteria,  their  presence  is  more  readily  detected  when 
they  are  stained.  Unfortunately,  however,  other  elements 
in  the  field  take  up  the  staining  material,  thus  rendering 
the  presence  of  bacteria  less  effective.  We  possess  a 
method  of  staining  which  has  the  distinct  virtue  of 
staining  only  bacteria,  leaving  pus  cells,  debris,  etc.,  intact 
— i.e.,  Gram's  method.  Moreover,  this  method  of  staining 
has  another  advantage,  for  by  it  some  bacteria  are  stained 
and  some  are  not.  It  is  therefore  used  as  a  diagnostic 
stain,  as  we  have  certain  bacteria  which  stain  by  Gram's 
method,  and  called  "  Gram-positive,"  and  other  bacteria 
which  do  not  stain — "  Gram-negative." 

Gram's  Method  of  Staining. 

Method. — Spread  the  fluid  on  a  slide,  dry,  and  fix  as 
already  described.  Stain  for  two  or  three  minutes  with 
carbol  gentian  violet  or  aniline  gentian  violet;  wash  in 
water  to  remove  excess  of  stain.  Flood  with  "  iodine  solu- 
tion "  (iodine,  1  part ;  pot.  iodide,  2  parts ;  water,  300  parts) 
for  about  one  minute ;  wash  off  in  alcohol  until  no  more  stain 
comes  away.  If  the  slide  is  left  very  pale,  one  may  usually 
take  it  for  granted  there  are  no  Gram-positive  bacteria 
present,  or  at  least  very  few.  The  film  should  now  be  sub- 
jected to  a  contrast  stain,  carbol  f  uchsin  (diluted)  or  neutral 
red ;  wash  in  tap-water,  dry  with  filter-paper,  mount,  and 
examine.  If  the  bacteria  are  Gram-positive  they  will  be 
dark  violet,  almost  black,  in  colour;  those  Gram-negative 
will  follow  the  contrast  stain,  as  will  blood-corpuscles, 
pus,  cells,  debris,  etc. 


VACCINE-THERAPY  29 

The  following  is  a  list  of  the  more  common  Gram  positive 
and  negative  bacteria : 

Gram-positive  : 

Staphylococcus  albus,  aureus,  citreus. 
Streptococcus. 
Pneumococcus,  diplococcus. 
Bacillus  of  anthrax. 
„        of  tuberculosis. 
„        of  diphtheria. 
„        of  tetanus. 
„        of  swine  erysipelas. 
„        of  blackquarter. 
„        of  acne. 
Micrococcus  tetragenus. 

actinomycosis. 

discomycosis. 

bothriomycosis. 

Gram-negative : 
Bacillus  coli. 

„        mallei. 

„        pyocyaneus. 
Bacillus  of  malignant  oedema. 

The  following  remarks  will  apply  only  to  the  most  useful 
and  common  stains  required  in  everyday  practice  : 

1.  Gentian  violet. 

2.  Methylene  violet. 

3.  Fuchsin. 

4.  Carbol-thionin. 

5.  Neutral  red. 

6.  Eosin. 

Any  of  these  stain  bacteria,  cells,  and  nuclei. 

Aniline  Gentian  Violet. 

Place  in  a  bottle  some  aniline  oil,  and  add  water  (always 
distilled)  ;  shake  well,  seeing  that  more  oil  is  added  than 


30  CLINICAL  BACTERIOLOGY  AND 

can  be  dissolved.  Allow  this  to  settle,  and  then  filter 
through  a  double  thickness  of  filter-paper.  To  9  parts  of 
this  solution  add  1  part  of  a  saturated  alcoholic  solution  of 
gentian  violet. 

This  stain  keeps  much  better  if  carbolic  lotion  1  in  40  is 
added. 

Loffler's  Methylene  Blue. 

This  stain  is  made  by  adding  30  c.c.  of  a  saturated  alco- 
holic solution  of  methylene  blue  to  100  c.c.  of  a  1  in  10,000 
solution  of  caustic  potash.  The  potash  solution  is  made  by 
taking  1  c.c.  of  a  10  per  cent,  solution  of  caustic  potash  and 
making  up  to  100  c.c.  with  water;  shake  well,  and  pour 
away  90  c.c,  making  up  the  remainder  to  100  c.c.  with 
water,  and  shaking  thoroughly.  Now  add  1  minim  of  the 
10  per  cent,  solution  to  2  ounces  of  water. 

This  stain  keeps  well. 

Carbol-Fuchsin  (Ziehl-Neelsen). 

This  stain  is  prepared  by  adding  a  saturated  alcoholic 
solution  of  fuchsin  to  carbolic  lotion  (1  in  20)  until  the 
fluid  has  lost  its  transparency,  the  preparation  usually 
being  about  1  in  9  strength. 

The  above  makes  a  very  powerful  stain,  and  should  be 
diluted,  when  required,  with  five  times  its  volume  of  distilled 
water. 

Carbol-Thionin. 

This  stain  can  be  used  instead  of  methylene  blue.     It 
is  made  by  adding  1  gramme  of  thionin  to  100  c.c.  of 
1  in  40  solution  of  carbolic  acid  in  distilled  water. 

Particular  care  should  be  taken  to  always  filter  this 
stain,  as  it  crystallizes  very  readily ;  in  fact,  it  is  a  good 
axiom  to  filter  all  stains  before  using  as  a  matter  of 
routine.  Before  use  dilute  with  equal  quantities  of  distilled 
water. 


VACCINE-THERAPY  31 

Eosin  and  Neutral  Red. 

These  stains  are  most  commonly  used  as  contrast  stains. 
They  are  particularly  clean  stains,  the  process  taking  place 
in  half  a  minute  to  one  minute. 

Capsule-Staining. 

Many  bacteria  are  invested  by  capsules,  and  their 
presence  or  absence  assists  in  their  identification.  To  stain 
the  capsule,  dip  the  specimen  in  a  mixture  of  equal  parts  of 
carbol-fuchsin  and  distilled  water  for  a  few  seconds,  rinse 
in  water,  stain  for  fifteen  seconds  in  an  aqueous  solution  of 
gentian  violet,  wash  in  water,  dry  on  filter-paper,  and 
mount. 

Flagella-Staining. 

Many  organisms  possess  delicate  plasmic  outgrowths, 
known  as  "  flagella,"  and  by  reason  of  their  delicate  con- 
struction they  are  somewhat  difficult  to  stain.  To  show 
them  to  the  best  advantage,  it  is  advisable  to  have  a  well- 
diluted  field  of  bacteria.  Take  a  very  small  quantity  of 
young  culture  and  place  it  on  a  slide.  Dilute  with  water. 
From  this  take  a  loopf  ul  and  spread  it  rapidly  on  another 
slide,  and  allow  it  to  air  dry;  then  fix  by  passing  it 
through  the  flame  twice,  taking  care  not  to  over-heat. 
Various  methods  are  adopted  for  staining  flagella. 

Pitfield's  Method  is  as  follows  : 

Take— 

(i.)   Saturated  solution  of  alum    ...  ...      10  c.c. 

Saturated  alcoholic  solution  of  gentian 

violet  1  c.c. 

Filter  and  bottle. 

(ii.)  Tannic  acid 1  gramme 

Distilled  water  10  c.c. 

Filter  and  bottle. 


32  CLINICAL  BACTERIOLOGY  AND 

Mix  an  equal  given  quantity  of  the  two  solutions  to- 
gether, and  flood  the  specimen  already  prepared  with  it. 
Hold  it  over  flame  of  spirit  lamp  until  it  nearly  boils.  Put 
it  aside  for  one  minute,  and  wash  in  water ;  dry,  stain  with 
aniline  gentian  violet  for  a  second  or  two,  wash  in  water, 
dry,  and  mount. 

Bonhill's  Method.  —  Take  a  small  quantity  of  culture 
material  on  a  platinum  loop,  and  move  it  about  in  a  little 
distilled  water  placed  in  a  test-tube.  Leave  the  tube  un- 
disturbed for  six  minutes,  and  then  place  1  drop  on  a  clean 
grease-free  cover-glass  and  dry  in  air.  Now  fix  in  the 
flame,  taking  care  not  to  overheat,  and  pour  some  orcein 
solution  on  a  watch-glass;  float  the  cover-glass  specimen 
side  downwards  on  the  surface  of  the  stain,  and  heat  gently, 
leaving  the  specimen  in  the  stain  for  fifteen  minutes.  Wash 
in  water  and  examine,  and  if  satisfactory  mount  in  balsam, 
but  if  not  re-stain. 

Spore-Staining. 

The  fact  that  spores  are  difficult  to  stain,  while  the  rest 
of  the  organism  is  not,  suggests  that  the  former  is  covered 
by  a  specially  thick  envelope,  which  prevents  the  ingress 
of  the  stain. 

Method  I. — Take  a  film  made  in  the  ordinary  way,  and 
flood  with  carbol-fuchsin  and  warm  for  fifteen  to  twenty 
minutes.  Wash  in  water,  and  dip  for  a  second  or  so  in 
1  per  cent,  solution  of  sulphuric  acid.  Wash  lightly,  and 
counter-stain  by  methylene  blue ;  wash,  dry,  and  mount. 

Method  II. — Moeller  stain  : 

Prepare  slide  in  the  ordinary  manner.  Keep  in  absolute 
alcohol  for  two  minutes,  and  then  in  chloroform.  Wash  in 
water,  allow  the  slide  to  stand  in  a  5  per  cent,  solution 
of  chromic  acid  for  two  minutes,  wash,  and  then  stain  with 
warm  carbol-fuchsin  for  ten  minutes.  Wash,  decolorize 
in  a  1  per  cent,  solution  of  sulphuric  acid.  Wash,  counter- 
stain  with  methylene  blue  for  one  minute.  Wash,  dry,  and 
mount. 


VACCINE-THERAPY  33 

Blood  Stains. 

The  two  in  most  common  use  are  Jenner's  and  Irish- 
man's, and  are  best  bought  already  prepared. 

Jenner's  Stain. — A  film  is  allowed  to  air  dry  and  then 
flooded  with  the  stain.  This  stain  fixes  as  well  as  stains 
the  specimen,  taking  three  minutes  to  act.  It  is  then  poured 
off,  washed  in  water,  dried,  and  mounted. 

Leishman's  Stain. — A  film  is  prepared  and  air  dried. 
The  specimen  is  flooded  with  the  stain  and  allowed  to  act 
for  thirty  seconds,  after  which  a  double  volume  of  distilled 
water  to  stain  is  added  on  to  the  film  and  mixed  with  a 
platinum  loop.  This  diluted  stain  is  now  allowed  to  act 
for  five  minutes.  Wash  in  water,  leaving  some  water  on 
the  film  for  half  a  minute  to  bring  out  the  colour.  Dry 
and  mount. 


CHAPTER  V 

IDENTIFICATION  OF  BACTERIA— GENERAL 
PRINCIPLES 

When  examining  morbid  material  or  blood  for  bacteria, 
it  does  not  follow  that  because  microscopical  examination 
fails  to  reveal  them  that  they  are  absent.  They  may  be 
so  few  in  number  as  to  escape  notice,  or  the  material 
examined  may  not  possess  any  bacteria  in  that  particular 
sample.  This  is  commonly  seen  in  tuberculosis  of  the 
udder,  where  the  milk  may  be  centrifugalized  and  yet  no 
bacilli  are  revealed.  Again,  in  abscess  formations  the 
digesting  influence  of  the  purulent  matter  may  be  so 
pronounced  that  the  active  organisms  are  only  to  be  found 
in  the  cyst  wall  or  its  neighbourhood.  Moreover,  it  is 
well  to  be  careful  in  forming  an  opinion  of  the  presence 
of  bacteria  taken  from  animal  discharges  after  dissolution, 
for  extraneous  contamination  rapidly  takes  place  from  the 
air  or  from  the  intestines.  In  the  case  of  an  animal  dying 
from  anthrax,  for  example,  we  know  hoAV  rapidly  the  blood 
becomes  contaminated  with  putrefactive  organisms  from 
the  intestines  after  death,  thereby  adding  greatly  to  the 
difficulty  of  arriving  at  an  accurate  diagnosis. 

Hanging- Drop  Preparations. 

Hanging -drop  preparations  are  used  principally  to 
identify  the  motility  or  otherwise  of  bacteria,  and  also 
to  observe  their  growth  and  development.     Hollow-ground 

34 


VACCINE-THERAPY 


35 


glass  slides  are  used,  the  circumference  being  painted  with 
immersion  oil,  or  a  wall  may  be  built  up  with  vaseline.  It 
is  most  important  not  to  place  too  large  a  drop  on  the 
cover-glass,  for  by  so  doing  it  will  reach  the  well  of  the 
slide,  and  the  fluid  will  run  by  capillarity,  the  hanging  drop 
thereby  disappearing.  Take  a  drop  of  emulsion  (bac- 
terial), well  diluted  with  broth  or  other  media,  and  place 
it  on  a  cover-glass ;  invert  the  hollow-ground  slide  over  it ; 
press  it  down  so  that  the  oil  round  the  well  adheres  to  the 
cover-glass  evenly.  Now  invert  the  slide,  and  the  hanging 
drop  is  complete  and  ready  for  examination.  Examine  with 
a  low-power  lens,  and  move  the  slide  about  when  the  edge 


® 

— —    ^M    \ 


A      B. 


Fig.  17. — Hanging- Drop  Preparation. 


of  the  hanging  drop  is  seen  in  the  field,  opening  and 
shutting  the  diaphragm  until  the  field  becomes  faintly 
illuminated.  Now  focus  a  little  deeper,  and  the  unstained 
bacteria  will  come  into  view,  and  note  if  motile  in  pairs  or 
clumps,  etc.  If  active  they  will  be  darting  about  in  all 
directions,  but  if  sluggish  they  can  be  revived  by  placing 
the  slide  in  an  incubator  for  a  short  time.  Needless  to 
add,  Brownian  movements  must  not  be  mistaken  for  natural 
bacillary  motion.  A  hanging-drop  preparation  from  cultures 
should  be  examined,  and  the  shape,  etc.,  of  organisms  noted, 
whether 

1.  Cocci  or  bacilli. 

2.  Motile  or  non-motile. 

3.  Grouping — clumps,  pairs,  chains,  etc. 


36  CLINICAL  BACTERIOLOGY  AND 

Three  films  should  then  be  made,  dried,  and  stained : 

1.  Carbol  methylene  blue. 

2.  Carbol-fuchsin  (Ziehl-Neelsen). 

3.  Gram's  method. 

Differential  Diagnosis  of  the   More  Common   Patho- 
genic Cocci. 

A  hanging-drop  specimen  should  be  made. 

A.  Non-Motile. — 1.  Staphylococci,  single  or  in  clumps. 
2.  Streptococci,  single,  pairs,  or  in  chains.  3.  Pneumo- 
cocci,  pairs  or  short  chains.  4.  Micrococcus  tetragenus, 
usually  in  fours.  5.  Micrococcus  catarrhalis,  single  or 
pairs. 

(a)  Grain-Positive. — (1)  Staphylococci;  (2)  streptococci; 
(3)  pneumococci;   (4)  tetragenus. 

Staphylococci. 
Biological  Characters. — 1.  In  broth   the   medium   be- 
comes turbid,  with  a  white,  yellow,  or  brown  layer  settling 
at  the  bottom. 

2.  On  agar  Staphylococcus  albus  forms  a  white,  elevated 
growth,  moist  and  glistening.  Staphylococcus  aureus  forms 
a  golden-yellow  growth.  Staphylococcus  citreus  forms  a 
lemon-coloured  growth. 

3.  In  gelatin  (stab  culture)  growth  takes  place  along 
the  whole  length  of  the  puncture,  followed  by  liquefaction 
in  two  or  three  days,  and  ending  in  a  deposit. 

Staining. — Methylene  blue  and  Gram's. 

Microscopical  Appearances. — Note  characteristic  bunches 
of  grape-like  formation. 

Facultative  Anaerobic. — Minimum  temperature  6°  C. ; 
optimum,  35°  C. ;  maximum,  45°  C. 

Streptococci. 
Biological  Characters. — 1.  In  broth  small  granules  de- 
velop, growing  sparingly. 


VACCINE-THERAPY  37 

2.  On  blood-agar  small  distinct  colonies  develop,  with  a 
clear  halo  round  them. 

3.  On  gelatin  small  white  granular  colonies  form. 
Gelatin  is  not  liquefied.  In  stab  culture  distinct  colonies 
form  along  the  track  of  the  needle. 

4.  On  potato  growth  very  sparing. 
Staining. — Carbol-methylene  blue  and  Gram's. 
Microscopical  Appearances. — Note  characteristic  long  or 

short  chain  formations. 

Facultative  Anaerobic. — Minimum  temperature,  10°  C.j 
optimum,  35°  C.  Growth  also  takes  place  at  room 
temperature. 

Pneumococci. 
Biological   Characters. — 1.  In  broth  difficult  to  grow, 
and  present  nothing  characteristic. 

2.  On  gelatin  growth  seldom  takes  place. 

3.  On  blood-agar  small  colonies  develop,  the  trans- 
parent medium  becoming  a  dirty  brown. 

Staining. — Carbol-methylene  blue  for  original  specimen; 
Gram's  stain  for  cultivated  specimen ;  and  also  stain 
capsule  with  a  capsule  stain. 

Microscopical  Appearances. — The  cocci  are  small,  lanceo- 
late in  shape,  and  arranged  in  pairs.  If  further  proof  is 
required,  inoculate  a  rabbit,  make  a  film  of  heart's  blood, 
and  note  capsule  by  capsular  stain. 

Aerobic. — Minimum  temperature,  20°  C;  optimum,  37°  C.; 
maximum,  40°  C. 

Micrococcus  Tetragenus. 
Biological    Characters. — 1.    In    gelatin    stab    a    thick 
growth  takes  place,  and  a  thick  white  disc  forms  on  the 
surface.     On  gelatin  plates  white  round  colonies  form. 

2.  On  agar  at  room  temperature  moist  white  colonies 
appear. 

3.  On  potato  a  thick,  irregular  slimy  patch  appears  at 
room  temperature. 


38  CLINICAL  BACTERIOLOGY  AND 

Staining. — For  original  pus  use  Welsh's  method.  After 
cultivation  use  carbol-methylene  blue  and  Gram's  method. 

Microscopical  Appearances. — Cocci  are  arranged  in  fours. 

Aerobic. — Minimum  temperature,  15°  0.;  optimum,  35°  C.j 
maximum,  40°  C. 

(b)   Gram-Negative. — Micrococcus  Catarrhalis* 

Biological  Characters. — 1.  On  gelatin  plates  at  room 
temperature,  20°  C,  the  growth  is  very  rapid  and  the 
colonies  tough. 

2.   On  agar  colonies  very  tough  and  opaque. 

Staining. — By  Gram's  counter-stain  neutral  red. 

Microscopical  Characteristics.  —  The  cocci  are  usually 
arranged  in  pairs. 

Aerobic. — Growing  at  room  temperature  freely.  Minimum 
temperature,  18°  C.  j  optimum,  37°  C. ;  maximum,  40°  C 

Differential  Diagnosis  of  the  More   Common  Patho- 
genic Bacilli. 

A  hanging-drop  specimen  should  be  made. 

A.  Motile. — I.  B.  tetani.  2.  B.  cedematus  maligni. 
3.  B.  pyocyaneus.  4.  B.  coli  communis.  5.  B.  anthracis 
symptomatica.     6.  B.  bronchosepticus  of  Ferry. 

(a)   Gram  Positive. — Bacillus  Tetani. 
Biological   Characters. — 1.    In    broth  anaerobically  the 
medium  becomes  clouded. 

2.  In  gelatin  stab  the  growth  radiates  into  the  medium, 
liquefies  slowly,  and  forms  gas. 

3.  On  agar  growths  form  giving  the  colonies  the  appear- 
ance of  very  fine  wool. 

4.  In  agar  stab  the  outgrowths  give  the  appearance  of  a 
fir-tree. 

*  A  microbe  having  close  characteristics  of  the  M.  catarrhalis  of 
man  has  been  isolated  by  the  author  from  catarrhal  discharges  of  horses 
suffering  from  influenza. 


VACCINE-THERAPY  39 

Staining  on  Tissue. — Carbol-methylene  blue  and  cultures 
by  Gram's  method. 

Microscopically  the  bacilli  when  speculating  are  like  drum- 
sticks, the  spores  being  at  the  ends. 

Anaerobic. — No  growth  at  20°  C.  On  broth  or  agar 
cultivation  at  37°  C.  in  anaerobic  tubes  after  subjecting 
the  original  material  to  80°  C.  for  thirty  minutes  to  kill 
vegetable  forms. 

(6)    Gram  Negative. — Bacillus  (Edematis  Maligni. 

Biological  Characters. — 1.  In  broth  becomes  cloudy. 

2.  On  gelatin  bright  grey  colonies  surrounded  by  spread- 
ing borders.  In  stab  culture  1  inch  below  the  surface  white 
side-branches  form. 

3.  On  agar  a  thick  network  of  threads  form. 
Staining. — Carbol-methylene  blue. 

Microscopical  Appearances.  —  The  bacilli  are  long  and 
slender  and  often  in  threads,  ends  rounded,  and  spores  are 
formed  in  the  middle. 

Strongly  anaerobic,  growing  well  at  room  or  incubator 
temperature. 

Bacillus  Pyocyaneus. 

Biological  Characters. — 1.  In  broth  the  growth  makes 
medium  cloudy. 

2.  On  gelatin  plates  flat,  irregular  colonies  form  with 
rapid  liquefaction,  the  whole  medium  becoming  green. 

3.  On  agar  the  colonies  form  a  white  upper  layer,  and  a 
deeper  layer  is  coloured  green. 

4.  On  potato  a  greenish-yellow  or  brown  layer  forms. 
Staining. — Carbol-methylene   blue  and   original  culture 

by  Gram's  method.     Contrast  stain  neutral  red. 

Microscopical  Appearances. — The  bacilli  are  often  arranged 
in  pairs,  and  form  filaments. 

Anaerobic,  growing  either  at  room  or  incubator  tem- 
perature. 


40  CLINICAL  BACTEKIOLOGY 

Bacillus  Coli  Communis. 
Biological  Characters. — 1.  In  broth  it  forms  a  diffused 
cloudiness  of  the  medium. 

2.  On  gelatin  -plates  it  forms  iridescent  colonies.  Gelatin 
does  not  liquefy. 

3.  In  gelatin  stab  culture  it  grows  in  the  form  of  a  nail. 

4.  On  agar  and  blood  serum  a  thick  layer  forms,  moist 
and  slimy. 

5.  On  potato  the  growth  at  20°  C.  is  brown  and  slimy. 

6.  In  litmus  milk  it  forms  acid,  gas,  and  clots. 
Staining.  —  Original    carbol  -  methylene    blue  ;    culture 

Gram's  method,  and  weak  carbol -fuchsin  as  a  contrast 
stain. 

Microscopical  Appearances. — Short  motile  rods,  mostly  in 
pairs,  sometimes  in  threads. 

Both  aerobic  and  anaerobic.  Grows  at  room  or  in- 
cubator temperature. 

Bacillus  Anthracis  Symptomatica. 
Biological    Characters.  —  1.     On    gelatin  plates   in   an 
atmosphere  of  hydrogen  a  radiating  dull  area  forms,  the 
gelatin  liquefies,  and  a  dark  lobulated  centre  develops. 

2.  In  deep  stab  glucose  gelatin  gas  forms. 

3.  In  glucose  agar  it  forms  a  dense  grey-coloured  growth, 
and  gives  off  a  pungent-smelling  gas. 

Staining. —  Carbol-methylene  blue,  but  not  ordinarily  by 
Gram's  method. 

Microscopical  Appearances. — The  bacillus  is  motile  with 
rounded  ends,  usually  single,  sometimes  forming  threads. 
Spores  form  at  the  ends  or  in  the  middle. 

Strictly  anaerobic,  growing  at  room  or  incubator  tem- 
perature. 

Bacillus  Bronchosepticus   [Ferry). 

Biological  Characters. — 1.  In  broth  cloudiness  take 
place  after  twenty-four  hours. 


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42 


CLINICAL  BACTERIOLOGY  AND 


2.  In  gelatin  (stab  culture)  after  twenty-four  hours  a  fili- 
form growth  occurs.     No  liquefaction  of  gelatin. 

3.  On  agar  a  moderate  growth,  slightly  raised,  surface 
moist  and  glistening,  of  a  sticky  consistence. 

4.  On  'potato  after  twenty-four  hours  growth  rather  abun- 
dant, surface  uneven,  moist  and  glistening,  raised  and  of  a 
light  tan  colour,  which  darkens  with  age. 


Fig.  20. — Surface  Elevation  of  Colonies. 

(1)  Flat.    (2)  Raised.    (3)  Convex.    (4)  Conical.    (5)  Umbilicate. 

5.  In  litmus  milk  after  five  days  colour  of  medium  at  top 
deep  blue,  with  a  brown  sediment  at  the  bottom. 

Staining. — Loeffier's  methylene  blue. 

Microscopical  Appearances. — A  short,  narrow  bacillus, 
single  or  in  pairs,  chains,  or  even  filaments. 

Aerobic. — Optimum  temperature,  37°  C. 


VACCINE-THERAPY  48 

B.    Non- Motile:    (a)    Gram-Positive. — Bacilli   of   acne, 
anthrax,  tuberculosis,  diphtheria,  and  actinomycosis.* 


Bacillus  Acne  Contagiosa  Equi. 

Biological  Characters. — 1.  In  gelatin  stab  white  colonies 
develop  along  the  course  of  the  needle. 

2.  On  agar  white  colonies  growing  very  slowly. 

Staining.— Difficult  to  stain  with  ordinary  dyes,  but  by 
Gram's  method. 

Microscopical  Appearances. — Small  ovoid  bacilli,  single, 
and  also  forming  chains. 

Anaerobic — at  least,  at  first.    Optimum  temperature,  37°  C 

Bacillus  Anthracis. 

Biological  Characters. — 1.  On  gelatin  plates  small  white 
colonies  appear,  and  if  examined  with  an  eyepiece,  charac- 
teristic threads  projecting  from  the  edges  of  the  colonies 
are  noted.  In  gelatin  stab  after  twenty-four  hours  a  thick 
white  central  column  appears,  from  which  radiate  lateral 
branches,  giving  the  whole  growth  a  triangular  appearance, 
with  the  apex  at  the  bottom. 

Liquefaction  begins  at  the  top,  extending  downwards. 

2.  On  agar  growth  is  similar  to  gelatin  plates. 

Staining. — Original  material  ordinary  aniline  dyes,  and 
by  Gram's  method. 

Note  McFadyean's  violet  colour  reaction  with  methylene 
blue. 

Microscopical  Appearances. — The  bacilli  appear  as  large 
rods,  and  often  in  threads.  In  stained  specimens  the  ends 
are  square  and  the  body  slightly  concave,  giving  it  the 
appearance,  if  in  thread  form,  of  a  bamboo  cane. 

Facultative  Aerobic. — Minimum  temperature,  10°  C; 
optimum,  35°  C. ;  maximum,  45°  C. 

*  Actinomycosis  is  not  a  bacillus,  but  for  convenience  is  described 
here. 


44  CLINICAL  BACTERIOLOGY  AND 

Bacillus  Tuberculosis. 

Biological  Characters. — 1.  In  broth  a  light  yellow  mass 
forms,  increasing  slowly,  the  medium  remaining  clear. 

2.  On  glycerine  agar  a  dry,  wrinkled,  brownish-yellow 
mass  forms. 

A  guinea-pig  should  be  inoculated  with  the  virus  for 
diagnostic  purposes  as  described  on  p.  183. 

Staining.  —  Ziehl-Neelsen's  method,  as  described  on 
p.  177. 

The  bacillus  belongs  to  the  acid-fast  group — i.e.,B.  Leprae, 
B.  Butyricus,  B.  Smegmatise,  bacillus  of  Johne's  disease, 
bacillus  of  Timothy-grass,  etc. 

Microscopical  Appearances. — This  bacillus  is  a  small  thin 
rod,  slightly  bent,  in  culture.  A  chain  of  four  or  of  eight 
may  be  noted. 

This  bacillus  is  aerobic  and  anaerobic.  Minimum  tem- 
perature, 27°  C.j  optimum,  37°  C;  maximum,  41°  C.  Growth 
slow :  about  six  weeks. 

Bacillus  Diphtherias. 
Biological  Characters. — 1.  In  glucose  peptone  broth  note 
formation  of  acid,  but  no  gas. 

2.  On  gelatin  plates  small  white  columns  develop,  and 
growth  is  very  slow,  without  liquefaction. 

3.  In  gelatin  (stab  culture)  at  20°  C.  surface  growth  takes 
place,  and  the  stab  growth  takes  the  form  of  a  nail. 

4.  On  agar  small  transparent  elevated  columns  occur. 
Growth  scanty. 

4.  On  Loeffler's  serum  large  white  colonies  grow. 

Staining. — Methylene  blue  or  weak  carbol-fuchsin,  and 
Gram's  method. 

Microscopical  Appearances. — The  bacilli  are  often  arranged 
in  clumps.     In  shape  and  size  they  vary  considerably. 

This  bacillus  is  aerobic  and  anaerobic.  Growth  takes 
place  between  minimum  temperature,  10°  C. ;  optimum, 
35°  C. ;  and  maximum,  48°  C. 


VACCINE-THERAPY  45 

Streptothrix  Actinomycosis  Boris. 
Biological  Characters. — 1.  On  broth  the  medium  is  not 
coloured,  but  grey  masses  settle  at  the  bottom. 

2.  On  gelatin  plates  in  a  week  a  yellow  growth  develops. 

3.  In  gelatin  stab  firm  outgrowths  take  place,  with  slight 
liquefaction. 

4.  On  agar  opaque  colonies  grow. 

5.  On  glycerine  agar,  at  37°  C,  in  three  or  four  days 
appears  a  yellow  growth,  tinged  with  red,  adhering  firmly 
to  the  medium. 

6.  On  potato  colonies  form  of  a  yellow  colour,  tinged  with 
red,  and  look  as  though  covered  with  fine  hair. 

Staining. — By  Gram's  method  and  methylene  violet. 

Microscopical  Appearances. — If  from  pus  note  filaments 
and  spores  (Gram-positive).  Clubs  round  periphery  of 
colony,  radiating  from  the  centre  (Gram-negative). 

It  is  anaerobic  and  facultatively  aerobic. 

(b)   Gram-Negative — Bacillus  Mallei. 
Biological  Characters. — 1.  In  broth  a  diffused  cloudiness 
takes  place,  ultimately  developing  into   a  tenacious  sedi- 
ment. 

2.  On  glycerine  agar  a  moist  opaque  layer  is  formed. 

3.  On  potato  a  characteristic  growth  at  37°  C.  takes  place, 
at  first  amber  in  colour,  and  later  developing  into  a  reddish- 
brown. 

Staining. — Young  cultures  stain  readily  with  the  ordinary 
aniline  dyes. 

Microscopical  Appearances. — The  bacillus  is  a  short  rod, 
usually  straight,  but  sometimes  curved.  They  are  usually 
single,  rarely  in  pairs  or  short  chains. 

Involution  forms  are  frequently  produced;  clubs,  filaments, 
and  even  branches  have  been  noted. 

This  is  a  facultative  bacillus  growing  with  or  without 
oxygen.  Minimum  temperature,  25°  C;  optimum,  37°  C. ; 
maximum,  42°  C. 


46  CLINICAL  BACTERIOLOGY 

Bacillus  Friedlander. 
Biological  Characters. — 1.  On  gelatin  plates  it  grows 
forming  small  porcelain-like  clusters  without  liquefaction. 

2.  In  gelatin  (stab  culture)  at  20°  C.  a  white  growth 
appears  along  the  centre,  showing  the  characteristic  nail- 
head  formation.     Gas  formation  on  shaking  gelatin. 

3.  On  agar  a  white  layer  is  formed. 

4.  On  potato  a  whitish-yellow  layer  develops,  with  gas 
formation. 

Staining.  —  Carbol-  methylene  blue  or  weak  carbol- 
fuchsin. 

Microscopical  Appearances. — Short  rods  with  rounded 
ends,  and  surrounded  by  a  distinct  capsule  under  special 
circumstances  of  growth. 

This  bacillus  is  aerobic  and  anaerobic,  growing  at  room  or 
incubator  temperature. 


CHAPTER  VI 

GLASS-WORK  REQUISITES  AND  HOW  TO 
MAKE   THEM 

How  to  make  a  Glass-Cutting  Knife. 

Take  an  ordinary  pocket-knife,  heat  the  blade  until  it 
becomes  white  hot  in  the  flame  of  a  blow  lamp,  and  im- 
mediately plunge  it  into  cold  water,  keeping  it  there  until 
it  has  become  quite  cold ;  the  edge  should  then  be  whetted 
on  a  rough  stone.  This  process  will  give  it  a  rough  cutting 
surface,  capable  of  scratching  ordinary  glass  tubing,  etc. 

When  the  rough  edge  wears  off,  it  can  always  be  renewed 
by  re  whetting  it  on  the  stone. 

Glass  Tubing. 

For  the  making  of  capillary  pipettes,  etc.,  tubing  of 
about  5  millimetres  external  diameter  is  usually  used.  The 
glass  should  be  of  the  best  kind  if  we  desire  to  make 
pipettes  uniform  in  shape  and  calibre,  and,  as  the  glass 
itself  is  quite  inexpensive,  there  is  no  need  to  use  inferior 
quality. 

Method  of  making  Capillary  Pipettes. 

Take  a  piece  of  glass  tubing,  cut  it  into  lengths  of  about 
9  centimetres.  This  is  done  by  holding  a  piece  of  glass 
tubing  in  the  right  hand  and  against  the  ball  of  the  thumb, 
while  the  same  hand  is  armed  with  the  glass-cutting  knife, 
the  edge  of  which  is  made  to  rest  firmly  upon  the  glass 
tube.  With  the  left  hand  the  tubing  is  now  rotated,  and 
47 


48  CLINICAL  BACTERIOLOGY  AND 

the  result  will  be  an  even  circular  scratch,  at  which  point, 
when  pressure  is  applied,  the  tubing  will  break,  leaving 
the  two  ends  clean-cut. 

The  proper  length  of  tubing  thus  made  should  be  taken 
up  and  rotated  by  the  right  and  left  hand  in  the  flame 
of  the  blow  lamp,  the  flame  being  in  the  middle  of  the 
tubing.  When  it  has  become  sufficiently  plastic  to  move 
about  freely,  it  should  be  removed  from  the  flame  and 
steadily  drawn  out  until  it  reaches  about  6  inches  in 
length.  The  middle  of  this  tubing  should  now  be  allowed 
to  touch  the  flame,  and  with  gentle  traction  the  ends  part, 
leaving  two  capillary  pipettes. 

Many  failures  will  follow  the  operator's  labours  at  the 


c 


^3 

Tar 

Fig.  21. — A  Simple  Capillary  Pipette. 

outset,  but,  as  already  stated,  glass  tubing  is  cheap,  and 
perseverance  will  bring  its  own  reward. 

To  reduce  the  risk  of  these  failures,  however,  the  follow- 
ing points  should  be  noted  : 

The  glass  should  be  uniformly  heated  from  start  to  finish, 
and  the  rotation  should  be  complete,  failing  which  one  side 
would  become  more  plastic  than  the  other,  and  the  resulting 
capillary  would  be  removed  from  the  central  axis  of  the 
tube. 

Again,  the  flame  throughout  does  not  give  off  the  same 
amount  of  heat;  therefore  it  is  wise  to  move  the  tube  up 
and  down  in  the  flame,  and  also  from  side  to  side.  Failing 
this  precaution,  we  shall  find  a  bulb  corresponding  to  the 
centre  of  the  flame,  with  capillary  taperings  on  either  side, 
due  to  the  fact  that  the  greatest  amount  of  heat  is  found 
at  the  sides  of  the  flame. 


VACCINE-THERAPY  49 

Throttled  Capillary  Pipettes. 

These,  by  reason  of  their  mechanism,  regulate  the  air 
transmitted  by  the  rubber  teat,  and  thereby  control  the 
movements  of  the  fluid  in  the  narrow  stem. 

Two  such  kinds  of  pipettes  are  in  common  use — i.e., 
(a)  where  the  throttle  is  in  the  distal  end  of  the  capillary 
stem ;  (b)  where  a  smaller  pipette  is  inserted  in  the  proximal 
end  of  the  ordinary  pipette. 

(a)  Take  an  ordinary  pipette  made  in  the  manner  already 
described,  and  hold  the  end  in  the  flame  of  an  ordinary 
wax  vesta  for  a  second  or  so,  and  then  draw  it  out  with  a 
sudden  jerk,  cut  off  the  end  with  a  glass  knife,  leaving  a 
very  fine  point. 


A    (L 


Fig.  22. — Throttled  Capillary  Pipettes. 

(b)  Take  an  ordinary  pipette,  and  insert  a  short  pipette  of 
smaller  calibre  in  the  proximal  end  of  it. 

The  smaller  pipette  is  throttled  in  the  manner  as  above 
described.  A  coating  of  sealing-wax  is  now  placed  round 
its  shoulder,  the  proximal  end  of  the  large  pipette  gently 
heated,  and  the  small  pipette  inserted,  the  wax  making  a 
tight-fitting  joint. 

Method  of  sealing  up  a  Test-Tube. 

When  it  is  desired  to  seal  a  test-tube — as,  for  example, 
after  making  a  bacterial  emulsion,  and  we  wish  to  break 
down  the  bacterial  bundles,  chains,  etc.,  by  shaking — it  is 
imperative  that  the  open  end  should  be  closed. — To  do  this 

4 


50  CLINICAL  BACTERIOLOGY  AND 

satisfactorily  one  requires  to  exercise  great  care,  and  the 
beginner  is  well  advised  to  practise  on  empty  test-tubes 
first.  Take  a  test-tube  and  rotate  in  a  small  flame  first, 
and,  when  thoroughly  hot,  rotation  should  go  on  in  a  full 
flame  until  the  sides  of  the  glass  begin  to  fall  in.  It 
should  now  be  drawn  out  into  a  narrow  stem,  and  as  soon 
as  it  has  become  cool  it  is  reheated  in  a  smaller  flame, 
drawn  out  into  a  thin  capillary  stem,  which  is  broken  off, 
set  aside  again  to  cool,  and  the  point  sealed  in  a  very  small 
jet,  candle  flame,  or  by  a  wax  vesta. 

When  one  is  dealing  with  a  fluid-containing  test-tube, 
greater  skill  and  care  is  required.  In  the  first  place,  the 
operator  must  make  sure  the  glass  at  the  point  the  flame 
touches  is  absolutely  dry  by  careful  heating.  Then  the 
flame  must  not  come  too  near  the  fluid,  or  it  will  boil  and 
crack  the  glass,  and  probably  injure  at  the  same  time  the 


5 


Fig.  23. — Drawn-Out  and  sealed  Test-Tube. 

chemical  qualities  of  the  fluid.  This  can  be  prevented,  of 
course,  by  holding  the  test-tube,  while  being  heated  in  the 
flame,  at  a  proper  angle,  and  not  heating  the  tube  too  near 
the  surface  of  the  fluid.  Of  course,  if  the  flame  is  applied 
too  near  the  proximal  end  of  the  tube,  the  glass  becomes  so 
hot  as  to  scorch  one's  fingers. 

When  there  is  an  excess  of  the  fluid  in  the  tube  neces- 
sitating this  condition,  it  is  wise  to  heat  the  mouth  of  the 
tube  to  red  heat,  holding  the  base  by  the  left  hand  and  by 
a  piece  of  glass  tubing  in  the  right  hand,  held  also  in  the 
flame  until  it  becomes  quite  hot.  Now  fix  the  tubing  to 
the  rim  of  the  test-tube,  first  at  one  point  and  then  at  the 
diagonally  opposed  point,  and  draw  the  whole  into  a 
narrow  stem.  When  cool,  put  the  stem  in  the  flame  and 
draw  the  two  ends  apart,  sealing  up  the  remaining  opening 
in  the  stem  by  applying  a  small  flame  to  it. 


VACCINE-THERAPY  51 

How  to  seal  One  End  of  a  Glass  Tube. 

When  we  desire  to  have  a  tube  smaller  than  a  test-tube 
which  will  be  found  useful  in  working  out  the  opsonic  power, 
etc.,  we  take  a  piece  of  pipette  tubing,  and  heat  it  in  the 
middle  in  the  usual  way,  drawing  it  out  to  make  a  capillary 
stem  (A).  This  will  give  us  the  makings  of  two  tubes  by 
breaking  the  stem  in  the  middle.  We  now  reduce  the  flame 
and  apply  it  to  the  shoulder,  rotating  the  tube  in  the  mean- 
time with  the  right  and  left  hands,  and  when  plastic  the 
stem  is  still  further  drawn  out  until  it  is  melted  through  (B) . 
Now  apply  a  full  flame  to  the  bottom  of  the  tube,  and  when 


Fig.  24. — Sealing  off  One  End  of  a  Glass  Tube. 

red  hot  blow  down  the  tube  (G).  This  will  make  the  distal 
end  quite  round,  and,  if  too  much  glass  is  not  left  on  the 
bottom,  the  heat  alone  will  be  sufficient  to  round  it  off 
nicely. 

How  to  make  Blood-Capsules  (Wright's). 

These  are  very  useful  for  the  examination  and  collection 
of  blood  to  obtain  the  serum  in  working  out  the  opsonic 
power  of  a  case. 

Take  a  piece  of  glass  tubing,  draw  out  into  a  capillary 
stem  (Fig.  25), 

Now  heat  the  barrel  in  the  middle,  draw  it  out  into  a  stem, 


52  CLINICAL  BACTERIOLOGY  AND 

and,  before  it  has  become  too  cool  to  lose  its  plasticity,  bend 
it  towards  the  body  of  the  tube  up  to  about  an  angle  of 
25  to  30  degrees  (Fig.  26). 

The  stem   should   now  be  burned   through  at  a  short 


Fig.  25.— Wright's  Blood-Capsule. 

distance  from  the  bend,  the  complete  capsule   being  the 
shape  of  Fig.  27. 

To  collect  blood  in  this  capsule,  care  should  be  taken  to 
see  both  ends  are  open.  If  the  blood  is  being  taken  from 
a  small  superficial  vein  in  the  horse,  after  opening  the  vein 


Fig.  26. — Weight's  Blood-Capsule. 


with  a  lancet,  the  bent  end  should  be  placed  in  the  blood- 
drop,  and  by  capillary  attraction  the  blood  will  siphon  into 
the  tube,  while  the  air  will  recede  from  the  straight  end. 
When  sufficient  blood  has  been  collected,  heat  the  straight 
end  in  a  small  flame,  and  seal  it  up.     As  the  imprisoned  hot 


Fig.  27. — Wright's  Blood-Capsule. 

air  at  this  end  cools,  it  will  contract,  and  the  blood  will  be 
withdrawn  from  the  bent  end,  which  in  turn  may  be  sealed. 
The  blood  will  now  clot  and  separate  out  the  serum.  This 
squeezing  out  of  the  serum  may  be  hastened  by  centri. 


VACCINE-THERAPY 


53 


fugalizing  the  capsule,  and  where  need  be  the  serum  may- 
be pipetted  off  into  a  small  tube  for  further  use. 

Slides,  and  how  to  make  Blood-Films. 

It  is  most  essential  that  glass  slides  should  be  free  from 
grease ;  this  is  done  by  boiling  in  lysol,  then  in  soda,  next 
in  strong  acid,  and  washing  out  in  spirit. 

If  a  slide  is  greasy,  it  is  obvious  one  cannot  obtain  a  per- 
fectly even  film ;  and  where  one  is  making  a  blood-count, 
for  reasons  of  accuracy  it  is  essential  that  the  objects  in 


Fig.  28. — Slide,  showing  Blood-Drop. 

the  field  should  be  evenly  distributed.  By  rubbing  the 
surface  of  the  slide  with  emery-paper,  a  mechanical  rough- 
ness is  produced,  which  facilitates  an  even  distribution  of 
the  elements  in  the  fluid. 

In  making  a  film,  it  is  necessary  to  ascertain  which  is  the 
concave  and  which  the  convex  side  of  the  slide,  for  it  is 
obvious,  to  get  a  good  spread,  we  must  place  our  film  on  the 
convex  surface.  To  find  out  which  is  which,  rotate  the 
slide  on  a  flat  smooth  surface.  On  one  face  it  will  not  spin 
(concave) ;  on  the  other  it  will  (convex) . 

To  make  a  blood-film,  place  a  drop  of  blood  as  blood, 


54  CLINICAL  BACTERIOLOGY 

or  mixed  with  emulsion  (bacterial),  which  we  wish  to 
standardize,  on  one  end  of  the  slide,  conveniently  at 
the  left-hand  border  (Fig.  28),  which  is  firmly  held 
between  the  thumb  and  finger  of  the  left  hand.  The 
end  of  the  spreader*  should  now  be  placed  in  the  blood- 
drop,  and  moved  backwards  and  forwards  and  from  side 
to  side  until  the  drop  is  spread  out  along  the  whole  length 
of  the  spreader's  edge.  Now  draw  the  spreader  steadily 
along  the  surface  of  the  slide ;  this  will  give  us  a  more  or 
less  perfectly  homogeneous  film,  composed  of  red  blood- 
corpuscles,  a  few  small  white  blood-cells,  and  blood-plasma. 
Many  of  the  leucocytes,  owing  to  their  size,  will  be  drawn 
from  the  field  by  the  spreader. 

The  film  should  now  be  allowed  to  air  dry,  fixed  with  a 
fixing  agent  or  a  fixing  stain,  and  stained. 

*  How  to  make  a  spreader,  see  p.  25. 


CHAPTER  VII 

THE  PROTECTIVE  ELEMENTS  OF  THE 
BLOOD,  WHICH  PROTECT  THE  ANIMAL 
BODY  FROM  PATHOGENIC  BACTERIA 

The  science  of  bacteriology  has  clearly  demonstrated  that 
by  far  the  largest  majority  of  organic  diseases  from  which 
animals  suffer  are  due  to  the  direct  or  indirect  influences  of 
specific  bacteria.  To  be  convinced  of  this  we  only  require 
to  look  through  the  long  roll  of  bacteria  which  have  been 
isolated  and  are  capable  of  producing  disease.  And, 
considering  the  serious  ravages  that  these  bacteria  have 
produced  in  the  past,  can  it  be  wondered  at  that  the 
resources  of  thinking  men  have  been  strained  to  the  utmost 
to  find  some  means  of  checking  their  destructive  processes? 
In  evolving  a  scheme  likely  to  satisfactorily  fulfil  its 
intended  purpose — namely,  the  treatment  of  disease — various 
methods  have  been  adopted,  but  the  one  which  concerns 
us  here — i.e.,  sero- vaccine  therapy — has  laid  open  a  vast 
field  for  research,  and  offers  to  those  who  pursue  its 
subtle  courses  rich  rewards  and  unthought-of  possi- 
bilities. To  the  general  practitioner  his  everyday  occupa- 
tion is  a  prophylactic  and  curative  one ;  and  whatever 
line  of  thought  or  school  of  teaching  he  follows,  to  be 
successful  he  must  always  be  subservient  to  Nature,  and 
the  more  he  keeps  in  touch  with  Nature  and  her  methods, 
and  in  his  healing  art  follows  her  example  and  emulates 
her  ways,  the  more  certain  will  be  his  successes.  Many 
centuries  ago  the  ancients  noticed  that  one  attack  of  a 
specific  disease  conferred  upon  the  subject  a  certain  protec- 
tion against  further  infection,  and  it  appears  the  first  attempt 
to  utilize  this  protective  principle    came   from   the    East, 


56  CLINICAL  BACTERIOLOGY  AND 

where  we  find  a  mild  attack  of  smallpox  was  produced 
by  inoculating  the  discharge  from  a  pustule  to  ward  off 
dangers  of  further  infection.  Following  upon  this,  William 
Jenner  discovered  that  vaccination  with  cowpox  virus  was 
equally  efficacious  as  a  preventive  against  smallpox  in 
man.     This  laid  the  foundation  of  vaccine-therapy. 

Succeeding  Jenner's  discovery,  the  actual  cause  of  certain 
specific  diseases  —  namely,  bacteria  —  was  demonstrated. 
Then  came  Koch's  work  upon  the  method  of  isolating  and 
identifying  bacteria. 

At  a  later  date  Metchnikoff  demonstrated  his  "  power  of 
resistance  theory,"  and  showed  the  action  of  the  leucocytes 
in  the  lower  animal  in  their  endeavours  to  devour  invading 
bacteria,  and  this  he  called  "  phagocytosis."  It  was 
further  proved,  however,  that  the  serum  exercised  some 
specific  influence  upon  bacteria,  and  it  was  shown  that  if 
the  blood-plasma  alone  was  taken  the  growth  of  bacteria 
was  inhibited,  and  on  the  supposition  that  there  existed  in 
the  plasma  some  protective  elements,  Buchner  gave  them 
the  name  of  "  alexins." 

Further,  it  was  demonstrated  that  an  animal  suffering 
from  an  infectious  disease,  and  recovering,  had  elaborated 
within  its  economy  various  substances  inimical  to  the  growth 
of  the  specific  bacteria  and  the  creation  of  their  products. 
These  substances  appear  to  be  of  two  distinct  kinds  : 
(1)  those  elaborated  consequent  upon  tissue  activities  and 
always  present,  called  non-specific  antibodies ;  (2)  those 
manufactured  in  response  to  an  infection,  and  named 
specific  antibodies. 

These  antibodies  vary  in  character  and  action,  and  at 
the  present  time  little  is  known  about  them. 

The  following  are  some  of  the  principal  antibodies : 

Antitoxins. 

These  are  specific  bodies  which  counteract  poisons,  or 
toxins,  manufactured  within  the  animal  body  by  disease- 
producing  organisms. 


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VACCINE-THERAPY  57 

In  1890  Behring  showed,  if  infinitesimal  quantities  of  the 
toxins  of  tetanus  were  injected  into  an  animal,  with  later 
succeeding  increased  doses,  the  time  would  arrive  when  that 
animal  would  tolerate  such  large  doses  of  tetanus  poison  with- 
out ill  effect,  and  Avhich,  if  injected  into  an  animal  whose 
serum  was  non-immunized,  would  cause  death  from  tetanus. 
Moreover,  if  the  serum  of  an  immunized  animal  were  mixed 
with  an  equivalent  amount  of  the  poison,  and  injected  into  a 
non-immune  animal,  no  ill-effect  would  follow.  Further, 
if  a  dose  of  immune  serum  was  administered  within  a  short 
period  of  time  to  an  animal  previously  inoculated  with  the 
tetanus  bacillus,  the  disease  would  not  develop.  This  fact  is 
seen  daily  in  the  use  of  antitetanic  serum  as  a  prophylactic. 
Behring  shortly  afterwards  showed  that  a  similar  condi- 
tion existed  with  regard  to  the  bacillus  of  diphtheria,  when, 
by  treating  an  animal  with  the  toxins  of  this  organism,  a 
serum  could  be  obtained  which  is  capable  of  possessing 
prophylactic  and  curative  effects  upon  this  disease  in 
man. 

The  toxins  manufactured  by  these  two  bacteria  (tetanus 
and  diphtheria)  are  known  as  exotoxins,  which  are  soluble, 
and  given  off  from  the  bacilli  when  grown  on  a  suitable 
culture  media — in  contradistinction  to  endotoxins,  which 
are  insoluble,  and  are  only  set  free  by  the  death  and  dis- 
integration of  the  bacteria  themselves,  such  as  staphylo- 
cocci, streptococci,  Bacillus  coli  communis,  etc. 

Various  theories  have  been  put  forward  from  time  to 
time  to  explain  the  production  of  antitoxins  by  body  cells, 
but  the  most  universally  accepted  now  is  Ehrlich's  cell 
nutrition  and  side-chain  theory. 

As  regards  cell  nutrition,  Ehrlich  believes  the  protoplasm 
is  made  up  of  many  molecules  showing  an  affinity  for 
a  large  variety  of  food  materials.  These  molecules  are 
composed  of  a  central  portion  surrounded  by  atomic  groups, 
which  unite  with  certain  food  molecules,  binding  them  to 
the  cell.  The  affinities  of  these  groups  vary  :  hence  they 
are  supposed  to   be   differently   constituted,  and  he  calls 


58  CLINICAL  BACTERIOLOGY  AND 

thein  side-chains  or  receptors.  A  side-chain  attached  to  a 
cell  may  join  to  itself  a  particle  of  oxygen,  fat,  carbo- 
hydrate, etc.,  and  so  take  part  in  the  nourishment  of  the 
cell.  In  the  same  manner  it  may  unite  with  a  molecule  of 
poison,  such  as  a  toxin.  Now,  this  toxin,  through  the 
medium  of  the  side-chain,  becomes  part  of  the  cell, 
and  if  strong  enough  will  poison  it  and  actually  produce 
death  of  the  cell  itself ;  or  it  may  cause  death  only  of  the 
side-chain,  in  which  case  the  latter  would  be  thrown  off 
and  a  new  one  formed  by  the  cell.  Following  upon  the 
destruction  of  the  side-chain,  several  more  chains  are 
produced  to  take  its  place,  with  the  result  that  the  cell 
cannot  retain  them  all,  and  they  consequently  become 
detached  and  are  cast  off  into  the  lymph  around  the  cell, 
and  eventually  reach  the  blood-stream,  constituting  free 
side-chains,  whose  function  now  appears  to  be  to  unite  with 
the  molecules  of  the  toxin  before  it  reaches  the  cell.  In 
this  manner  these  free  side-chains  show  antitoxin  properties 
which  prevent  the  action  of  the  poison  upon  the  cells, 
giving  thereby  to  the  patient  an  immunity.  Further,  if  the 
serum  containing  these  free  side-chains  is  injected  into 
another  animal,  they  will  confer  upon  that  animal  the  same 
degree  of  immunity  as  was  possessed  by  the  original 
immunized  animal,  which  explains  the  prophylactic  and 
curative  action  of  antitoxin. 

Toxins  are  easily  destroyed  by  heat,  chemicals,  and  light, 
but  the  loss  of  toxicity  does  not  result  from  a  complete 
destruction  of  the  toxin  molecule,  for  it  is  still  able  to 
unite  with  the  antitoxin.  It  is  clear,  then,  that  the  toxin 
molecule  is  made  up  of  two  parts :  a  thermostable  portion, 
which  is  capable  of  uniting  with  cell  receptors  either  in  the 
cell  itself  or  free  as  antitoxin,  and  called  the  haptophore ; 
and  a  thermolabile  group,  which  causes  the  cell  injury  after 
union  by  means  of  the  haptophore  has  taken  place,  and 
called  the  toxophore.  When  the  toxophore  group  of  a 
toxin  has  been  destroyed,  that  which  remains  is  called  the 
toxoid. 


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59 


Agglutinins. 

Gruber  and  Durham  have  shown,  when  the  serum  of  an 
animal  suffering  from  a  bacterial  disease  is  added  to  a 
culture  of  the  specific  bacterium  of  that  disease,  they  col- 
lect together  in  masses,  leaving  the  rest  of  the  fluid  free 
from  their  presence.  It  is  supposed  they  (agglutinins)  are 
formed  in  lymphoid  tissue,  marrow,  and  the  spleen,  while 
Metchnikoff  noticed  the  peritoneal  exudate  was  even  richer 
than  the  blood  itself  in  agglutinins. 

Strangely  enough,  the  agglutination  of  bacteria  seems  to 
protect  them  from  the  death-producing  action  of  another 
antibody— i.e.,  lysin ;  and  if  that  is  so,  Nature  would  appear 
to  be  working  against  her  own  interests  and  defeating  her 


Fig.  29. — Agglutination  of  Bacteria  (Jowett) 

A,  Bacteria  evenly  distributed  ;  B,  Bacteria  agglutinated  by 


own  ends — a  very  improbable  presumption.  Allen  is  of 
opinion,  however,  the  bacteria  are  gathered  into  clumps  in 
the  manner  just  described,  so  that  they  may  be  caught  up 
in  the  various  tissues,  such  as  the  liver,  the  spleen,  and 
lymphatics,  where  the  immunizing  bodies  are  generated,  and 
their  destruction  thereby  brought  about. 

Widal  first  suggested  using  this  agglutinating  power  for 
diagnostic  purposes,  and  the  test  now  bears  his  name.  It 
was  found  that  many  kinds  of  micro-organisms  were  clumped 
by  the  serum  of  animals  immunized  against  them.     It  was 


60  CLINICAL  BACTERIOLOGY  AND 

thought  at  first  the  serum  taken  from  a  patient  suffering 
from  a  specific  disease  had  the  specific  agglutinating  power 
only,  but  later  it  was  demonstrated  that  the  undiluted  serum 
of  normal  persons  was  capable  of  producing  the  same  effect. 
It  was  then  decided  to  dilute  the  serum.  Thus,  one  loopful 
of  serum  was  mixed  with  nine  loopfuls  of  a  young  fresh 
bacterial  emulsion.  Then  the  strength  was  made  1  in  50, 
and  when  the  patient  was  suffering  from  the  specific  dis- 
ease the  agglutinating  power  would  still  be  in  evidence. 
The  length  of  time  allowed  for  agglutination  to  take  place 
is  usually  half  an  hour. 

Widal's  Test  for  Diagnostic  Purposes. 

Collect  the  blood  from  the  patient  in  a  Wright's  capsule. 
Allow  the  clot  to  shrink,  to  squeeze  out  the  serum ;  or,  to 
hasten  the  process,  centrifugalize  it,  placing  the  capsule  in 
the  centrifuge,  having,  of  course,  previously  sealed  the 
straight  end  of  the  capsule.  In  this  way  we  obtain  our 
serum.  We  now  take  a  young  culture,  preferably  grown 
on  broth,  make  a  hanging  drop,  microscopically  examine 
the  specimen,  and  note  the  disposition  and  motility  of  the 
bacteria. 

We  then  take  a  capillary  pipette,  make  a  unit  mark  with 
the  grease  pencil  on  the  stem  high  enough  up  to  contain 
1  to  2  cm.,  aspirate  one  unit  of  serum,  a  bubble  of  air,  one 
unit  volume  of  0-86  per  cent,  saline  solution,  a  bubble  of 
air,  one  volume  of  saline  solution,  and  so  on,  until  we  have 
taken  up  fourteen  volumes  of  the  solution,  each  separated  by 
a  column  of  air.  The  whole  is  now  expelled  from  the  pipette, 
and  again  aspirated  to  thoroughly  mix,  and  further  dilu- 
tion of  this  dilution  is  made — usually  three  dilutions, 
1  in  15,  1  in  25,  1  in  50. 

Three  hanging-drop  slides  are  next  prepared.  One  loopful 
of  the  diluted  serum  should  be  placed  on  three  cover-glasses 
(1  in  15,  1  in  25,  1  in  50),  and  to  each  is  added  and  mixed 
one  loopful  of  broth  culture  containing  the  organism.  The 
hanging    drops    are    now    examined     under    the    micro- 


VACCINE-THERAPY  6 1 

scope,  and  note  taken  of  the  disposition  of  the  bacteria  up 
to  an  hour,  but  usually  half  an  hour  is  long  enough.  If 
the  reaction  is  a  positive  one,  the  bacteria,  if  motile,  will 
lose  their  motility,  and  collect  themselves  into  clumps — 
agglutinate.  On  the  contrary,  if  the  reaction  is  a  negative 
one  no  clumping  will  take  place. 

Another  method  of  making  use  of  the  agglutinative 
reaction  for  diagnostic  purposes  is  to  add  a  measured 
volume  of  serum  to  a  known  quantity  of  the  culture  media 
in  a  test-tube.  In  due  course,  if  the  reaction  is  a  positive 
one,  a  white  precipitate  will  form  and  settle  on  the  bottom 
of  the  tube.  This  is  known  as  the  "  precipitation  test,"  and 
has  the  advantage  of  being  very  simple  in  its  application. 

Bacteriolysins. 

Under  this  heading,  we  find  there  exists  in  the  blood 
protective  bodies  capable  of  destroying  bacteria  by  solu- 
tion— bacteriolysins — and  the  process  is  known  as  "bac- 
teriolysis." It  appears  that  the  serum  of  the  blood  soon 
loses  its  bacteriolytic  properties  when  separated  from  the 
tissues. 

Experiments  have  proved  that  the  solution  of  the 
microbes  is  brought  about  by  the  interaction  of  at  least 
two  substances,  one  of  which  is  present  in  all  serums  in 
the  living  body,  but  disappears  on  heating  and  on  keeping 
at  room  temperature  in  from  five  to  eight  days.  The 
other  is  more  stable,  and  is  produced  during  the  process 
of  inoculation.  The  former  unstable  body,  found  in  all 
animals,  is  termed  "complement";  the  latter,  produced 
in  the  processes  of  immunity,  is  called  the  "  amboceptor." 

It  is  presumed  the  amboceptor  links  the  complement  to 
the  bacterium,  but  the  complement  remains  free  if  the 
suitable  amboceptor  is  not  present,  and  bacteriolysis  does 
not  take  place  (Ehrlich)  (Fig.  30). 

It  is  upon  this  process  of  bacteriolysis  that  Pfeiffer's 
reaction  depends,  which  is  useful  in  recognizing  the  exact 
species  of  bacteria. 


62  CLINICAL  BACTERIOLOGY  AND 

Method. — Take  a  mixture  of  the  bacterial  emulsion 
and  mix  with  it  a  small  quantity  of  serum  from  an 
animal  immune  to  the  bacterium  in  question,  and  inject 
into  the  peritoneal  cavity  of  a  healthy  guinea-pig.  After 
half  an  hour  the  peritoneal  fluid  and  exudate  is  micro- 
scopically examined;  and  if  the  reaction  is  a  positive  one 
the  organisms  will  be  found  to  have  undergone  degenera- 
tion, in  which  case  the  organism  will  be  found  to  belong  to 
the  same  species  as  the  one  which  the  already  immunized 
animal  was  invaded  by.     On  the  other  hand,  if  the  reaction 


Fig,  30. — A,  Micro-organism  ;  B,  Immune  Body  or  .Amboceptor; 
C,  Complement  (Ehrlich). 

is  a  negative  one  the  organism  belongs  to  a  different  species 
to  that  by  which  the  animal  was  immunized. 

Opsonins. 

Wright  has  shown  there  exists  in  blood-serum  an  anti- 
body which  possesses  the  specific  virtue  of  sensitizing  the 
bacteria  present,  and  in  this  manner  prepares  them  for 
the  devouring  activities  of  the  leucocytes.  He  calls  this 
antibody  "  opsonin,"  or  "  feast-preparer."  It  appears  this 
opsonic  power  which  the  serum  exerts  upon  each  kind  of 
bacterium  is  a   specific   one,  for  we  find  if  an  animal  is 


VACCINE-THERAPY  63 

suffering  from  a  bacterial  infection  and  his  opsonic  index 
to  that  infection  stand  at  a  certain  point  a  further  invasion 
by  another  specific  bacterium  need  not  alter  that  index  one 
way  or  another. 

In  estimating  the  opsonic  power,  we  ascertain  the  follow- 
ing valuable  points :  (1)  what  natural  resources  the  animal 
possesses — in  other  words,  at  what  point  his  index  stands ; 
(2)  during  the  progress  of  the  disease  we  can  note  whether 
that  index  rises  or  falls — in  short,  what  efforts  Nature  her- 
self is  displaying  in  her  endeavours  to  shake  off  the  in- 
vasion; (3)  where  we  are  adopting  vaccine-therapy,  the 
index  will  assist  us  in  forming  an  opinion  regarding  the 
suitability  of  the  dose,  and  to  what  degree  the  patient 
responds  to  our  vaccine  stimulus. 

Of  course,  as  we  shall  see  later,  clinical  observations  on 
these  three  points  also  are  of  great  assistance  to  us  in 
forming  our  conclusions  upon  the  various  phenomena  and 
immunizing  responses  made  manifest  during  the  progress 
of  a  specific  disease.  Although  in  practice  the  taking 
of  the  opsonic  power  is  not  essential  to  success,  it  gives 
us  a  very  valuable  and  very  delicate  guide  to  follow,  and 
well  repays  the  practitioner  who  will  take  the  extra  trouble 
to  work  it  out. 

The  Usual  Method  adopted  in  estimating  the  Opsonic 
Power  of  the  Blood. 

1.  Take  a  Wright's  blood-capsule,  and,  after  sterilizing 
the  skin  over  one  of  the  facial  veins  of  an  animal  belonging 
to  the  same  species  as  the  patient,  puncture  the  vessel  with 
a  small  lancet,  and  collect  the  blood  in  the  capsule.  In 
like  manner  collect  the  blood  from  the  patient  at  or  nearly 
at  the  same  time,  since  opsonin  after  a  length  of  time 
becomes  inert. 

2.  Take  the  bacterial  growth  we  desire  to  test,  and  not 
more  than  twenty-four  hours  old.  Add  1  or  2  c.c.  of  normal 
saline  solution  to  the  culture-tube ;  shake  gently  to  wash  off 
the  colonies.    The  resultant  emulsion  should  be  pipetted  off 


64  CLINICAL  BACTERIOLOGY  AND 

into  a  small  tube,  centrifugalized  for  a  few  minutes  to  assist 
in  breaking  down  the  clumps,  chains,  etc.,  and  then  well 
shaken,  after  sealing  the  tube,  as  described  on  p.  51,  Fig.  24. 
This  emulsion  should  be  of  a  faint  opalescent  colour. 

If  too  many  bacteria  are  in  emulsion,  it  may  be  almost 
white,  in  which  case  it  is  too  rich,  and  the  leucocytes  will 
take  up  an  excessive  number  of  bacteria,  which  will  render 
the  field  difficult  to  count.  Such  an  emulsion  should  be 
further  diluted. 

3.  Prepare  an  emulsion  of  living  leucocytes  in  the 
following  manner :  Take  about  10  c.c.  of  normal  saline 
solution,  and  add  ^  per  cent,  sodimn  citrate  solution  to 
prevent  coagulation,  of  the  blood  ;*  place  this  in  a  centri- 
fugalizing  tube  and  tvarm  to  blood  heat.  Then  prick  the 
facial  vein  of  the  healthy  animal  and  drop  into  the  tube 
1  or  2  c.c.  of  blood.  This  is  now  put  into  the  centrifuge 
and  centrifugalized  about  fifteen  minutes,  until  all  the 
corpuscles  have  gone  to  the  bottom  and  the  supernatant 
liquid  is  left  clear. 

The  deposit  will  show  the  red  corpuscles  at  the  bottom, 
and  on  the  top  a  thin  grey  layer  of  leucocytes.  Now 
aspirate  the  clear  fluid  with  a  capillary  pipette  and  teat, 
taking  care  not  to  disturb  the  layer  of  leucocytes. 

This  done,  suck  up  the  leucocytic  layer  and  some  of  the 
red  corpuscles,  and  put  them  in  a  small  tube,  thoroughly 
mixing  them  with  the  pipette,  giving  an  emulsion  of 
living  leucocytes  and  a  few  red  corpuscles. 

4.  Take  two  Wright's  pipettes,  with  rubber  teats,  and 
with  a  grease  pencil  mark  the  glass  stem  at  its  distal  end 
^  to  1  inch  from  the  point.  Fluid  taken  up  to  this  mark 
constitutes  one  unit  volume. 

We  should  now  have  in  front  of  us  on  the  laboratory 
table  a  slab  of  plasticine,  fixed  into  which  are  (a)  a  small 

*  Burroughs  Wellcome  and  Co.  make   convenient  soloids   of  the 
following  formula : 

Sodii  chloridi     ...  ...  ...  ...     0-075  gm. 

Sodii  citratis      ...  ...  ...  ...     0-05    gm. 


VACCINE-THERAPY  65 

tubule  each  containing  the  patient's  serum ;  (b)  control 
serum;  (c)  the  washed  emulsion  of  leucocytes;  (d)  the 
emulsion  of  bacteria. 

We  then  take  up  a  pipette  and  immerse  the  point  into 
the  emulsion  (bacterial),  and  draw  up  by  relaxing  the 
rubber  teat  one  volume — i.e.,  up  to  the  grease  pencil  mark 
— withdraw  the  point  from  the  emulsion,  and  slightly 
relaxing  the  pressure  upon  the  teat,  allow  the  ingress  of  a 
column  of  air.  In  like  manner  take  up  one  volume  of 
leucocytic  emulsion,  and  then  a  column  of  air.  Dip  the 
pipette  into  the  leucocytic  emulsion  again  and  take  up 
another  volume,  then  a  column  of  air,  and  lastly  draw  up 


D 


Fig.  31.-^4,  Pipette  with  Rubber  Teat  containing  in  Order 
the  Individual  Elements  separated  by  Columns  of  Air; 
B,  Pipette  showing  the  Elements  thoroughly  mixed  and 
ready  for  incubating. 

one  volume  of  serum.  In  the  pipette  we  now  have,  count- 
ing from  the  point  to  the  teat,  a  volume  of  serum,  a  column 
of  air,  a  volume  of  leucocytes,  a  column  of  air,  a  volume  of 
leucocytes,  a  column  of  air,  a  volume  of  emulsion.  The 
operator  having  laid  out  a  clean  slide  in  front  of  him,  the 
point  of  the  pipette  is  made  to  rest  upon  it,  and  the  fluid  in 
the  stem  expelled  and  sucked  up  several  times  to  thoroughly 
mix  the  contents. 

It  may  be  well  to  note  here  that  the  teat  should  perfectly 
fit  the  stem  of  the  glass,  failing  which  we  will  not  obtain 
the  same  free  aspirating  and  expelling  powers  we  should 
have.  This  point  can  be  proved  by  pressing  the  teat,  and 
then  sealing  the  end.     If  the  union  is  complete,  the  teat 

5 


66  CLINICAL  BACTERIOLOGY  AND 

will  remain  compressed;  if  incomplete,  it  will  gradually 
become  inflated. 

Again,  beginners  may  find  a  difficulty  in  controlling  the 
fluid  movements  in  the  stem  of  the  pipette.  This  with  prac- 
tice will  soon  be  overcome,  when  the  operator  acquires  that 
delicacy  of  touch  gained  by  practical  experience,  pro- 
vided, of  course,  the  end  of  the  pipette  has  been  properly 
drawn  out  and  squarely  cut. 

When  we  expel  the  contents  of  the  pipette  containing 
air-bubbles,  of  course  we  find  the  air-bubble  question 
troublesome  when  we  begin  to  aspirate.  To  obviate  this 
condition,  raise  the  glass  slide,  rest  the  end  of  the  pipette 
upon  it,  squeeze  the  teat,  and  allow  the  fluid  to  run  away 
from  the  point. 

When  bubbles  are  in  existence  on  the  slide,  we  aspirate 
from  the  far  side  of  the  drop;  in  this  manner  making  a 
kind  of  strainer. 

The  contents  having  been  thoroughly  mixed,  we  now 
aspirate  the  whole  fluid ;  suck  in  a  column  of  air  at  the 
point,  and  seal  the  end  in  the  flame. 

The  barrel  of  the  pipette  should  be  marked  with  the 
grease  pencil  with  the  owner's  name  or  number  of  patient, 
and  placed  in  the  incubator  at  37°  C.  for  fifteen  minutes, 
making  careful  note  of  the  time. 

Pipette  No.  2  should  be  treated  in  exactly  the  same 
way,  save  that  the  control  serum  takes  the  place  of  the 
patient's  serum,  and  incubated  for  fifteen  minutes  exactly. 

When  the  pipette  has  been  incubated  the  proper  time,  it 
is  taken  out.  A  teat  is  fixed  on  to  the  barrel,  the  pointed 
or  distal  end  of  the  stem  is  scratched  and  broken  off,  the 
whole  of  the  contents  expelled  on  to  a  slide,  and  thoroughly 
well  mixed,  as  previously  described. 

One  or  two  drops  should  now  be  placed  upon  a  clean, 
grease-free,  emery-paper-rubbed  slide  (on  its  convex  face), 
and  the  film  made  with  a  spreader.  It  is  well  to  make 
three  slides  in  this  manner,  so  that  one  may  choose  the 
best  film  in  the  end,  and  air  dried.     The  application  of 


VACCINE-THERAPY  67 

heat  to  fix  these  films  is  scarcely  practicable,  unless  in 
highly  skilled  hands.  As  fixing  agents  we  therefore  use 
formalin  or  corrosive  sublimate.  The  latter  is  probably  the 
most  useful,  speaking  generally. 

The  slide  is  placed  in  the  slide-rack  containing  a  satu- 
rated aqueous  solution  of  hyd.  perchlor.  for  half  to  one 
minute.  It  is  then  washed  under  the  tap,  air  dried,  and 
stained  with  carbol-thionin,  but  in  the  case  of  tubercle  by 
carbol-fuchsin  (hot). 

The  best  film  of  each  should  now  be  examined  by  a  low 
power,  to  ascertain  which  is  the  best,  and  then  by  a  -^  oil- 
immersion  lens,  and  many  of  the  polynuclear  leucocytes 
will  be  found  to  contain  bacteria. 

A  pencil  and  a  piece  of  paper  should  then  be  placed 
at  the  side  of  the  microscope,  and  a  count  of  the  leucocytes 
containing  bacteria  made  up  to  fifty,  first,  from  the  slide 
with  the  control  serum,  secondly,  from  the  slide  with  the 
patient's  serum,  the  ratio  between  the  two  giving  the 
opsonic  index. 

Thus,  if  the  control  serum  film  showed  that  the  50 
leucocytes  had  ingested  150  bacteria,  and  the  patients' 
serum  film  showed  that  the  50  leucocytes  had  ingested 
80  bacteria,  the  ratio  80^-150  =  0*53  gives  the  opsonic 
power  of  the  patient;  or,  assuming  the  control  animal's 
opsonic  power  to  be  normal,  the  patient's  would  be  nearly 
50  per  cent,  below  normal. 

In  such  a  condition  an  animal  is  a  bad  subject  to  resist 
bacterial  invasion.  It  has  been  shown  that  a  patient  may 
have  a  very  high  opsonic  index  against  a  bacterial  infection 
at  a  certain  stage  of  the  disease,  and  yet  die.  This  state  is- 
usually  explained  by  reason  of  the  fact  that  secondary  in- 
fection has  taken  place,  and  secondary  bacteria  are  the 
cause  of  dissolution. 

A  very  low  opsonic  power  suggests  either  that  the  animal 
is  suffering  from  the  direct  effects  of  a  specific  infection,  or 
its  resisting  powers  are  so  low  as  to  make  it  susceptible  to 
infection  should  opportunities  occur.    This  explains  largely 


68  CLINICAL  BACTERIOLOGY 

why  some  animals  are  more  predisposed  to  specific  infec- 
tions than  others. 

But,  of  course,  it  must  be  pointed  out  that  a  high  opsonic 
power  does  not  necessarily  mean  absolute  immunity  against 
bacterial  invasion,  for,  as  we  have  already  seen,  there  are 
other  protective  elements  in  the  blood,  and  in  addition  the 
living  tissue  and  organic  structures  of  the  body,  which 
play  their  part  in  the  role  of  conferring  and  sustaining 
immunity,  and  to  what  extent  each  can  act  by  itself  is  at 
present  not  known. 

Phagocytosis. 

The  first  important  theory  of  disease  resistance  was  put 
forward  by  Metchnikofr,  who  studied  the  behaviour  of  the 
leucocytes  in  the  lower  animals,  and  attributed  the  destruc- 
tion of  bacteria  to  their  activities.  Metchnikoff's  theory 
is  that  the  phagocytes  approach  and  devour  invading 
bacteria,  and  if  they  succeed  in  doing  this  the  disease  is 
checked  and  prevented.  It  has  been  already  pointed  out 
that  the  leucocytes  are  not  in  themselves  capable  of  acting 
in  the  capacity  of  invaders,  but,  as  Wright  and  others 
believe,  the  bacteria  must  be  in  some  manner  prepared  for 
them  before  digestion  can  take  place.  On  the  face  of  this 
it  appears  only  reasonable  to  suppose  that  phagocytosis  is 
only  one  link  of  the  complex  chain  which  every  living 
being  is  daily  forging  within  his  economy  in  his  fight 
against  more  or  less  constant  bacterial  invasions ;  and  it  is 
to  the  harmonious  working  of  all  these  protective  forces, 
presenting  a  strong,  vigorous  front,  that  we  owe  and  main- 
tain our  existence. 


CHAPTER  VIII 

VACCINES  AND  THEIR  MODE  OF 
PREPARATION 

From  the  previous  chapter  it  will  be  gathered  that  within 
the  animal  body  certain  protective  forces  exist,  which, 
should  a  bacterial  invasion  occur,  become  offensive  in  their 
action,  and  wage  war  against  the  invading  germs.  This 
warfare  is  taking  place  daily  in  every  human  and  animal 
body.  Sometimes  these  antibodies  are  so  strong  and  active 
that  complete  protection  takes  place,  and  no  disturbance 
is  made  manifest.  At  other  times  the  cardinal  symptoms 
of  disease  present  themselves,  and  after  running  their 
course  recovery  takes  place,  and  the  leucocytes  and  anti- 
bodies are  thereby  fortified.  In  other  cases  the  bacteria  or 
their  products  disintegrate,  and  destroy  Nature's  protective 
antibodies,  with  consequent  dissolution  of  the  animal. 

Medicinal  preparations  have  long  held  sway  in  com- 
bating disease,  but  how  many  practitioners  of  medicine  are 
there  who  are  absolutely  satisfied  with  the  curative  values 
of  drugs  in  specific  bacterial  diseases  ?  Nay,  more,  the 
longer  they  are  prescribed,  the  more  apparent  do  their 
limitations  become. 

Vaccine-therapy  not  only  carries  the  therapeutist  beyond 
this  limited  field,  but  in  many  cases  he  supplants  the 
medical  therapeutist  altogether,  and  that,  too,  often  to  the 
great  advantage  of  the  patient.  Moreover,  the  logic  of 
vaccine-therapy  is  based  upon  a  sound  reasoning  founda- 
tion which  has  the  advantage  of  imitating  Nature, 
and  endeavouring  to  effect  a  cure  in  the  same  way  as 
Nature  herself ;  and  in  those  cases  where  Nature,  usually 


70  CLINICAL  BACTERIOLOGY  AND 

through  a  conglomeration  of  circumstances,  fails  to  rise  to 
the  occasion,  the  immunizer  should  step  in,  and  with  his 
vaccines  supply  the  essential  stimulus  which  is  needed  to 
produce  the  various  antibodies  which  she  requires  to  battle 
against  bacterial  invasions. 

Some  animals  are  born  with  a  lesser  degree  of  resistance 
to  a  bacterial  infection  than  others;  and  also  this  dimin- 
ished resistance  is  particularly  noticeable  in  certain  strains 
of  families,  and  this  fact  often  led  the  older  practitioners 
astray,  when,  by  reason  of  the  persistent  recurrence  of  the 
disease  in  several  succeeding  generations,  they  insisted  that 
the  condition  must  be  hereditary,  when  there  was  nothing 
more  inherited  than  a  weakened  resistance.  It  is  in  cases 
such  as  these  that  vaccine-therapy  can  do  so  much.  With 
this  as  with  all  other  systems  of  treatment,  an  orthodox 
line  laid  down  and  dogmatically  followed  is  almost  certain 
to  lead  to  failure  and  mistrust,  and  it  is  probable  that  there 
is  no  other  system  of  medicine  which  requires  more  fore- 
thought, more  precision,  and  more  careful  analytical  study, 
on  the  part  of  the  immunizer,  than  vaccine-therapy.  For 
example,  an  animal  may  develop  strangles,  the  disease 
becomes  pysemic,  and  in  due  course  we  clear  the  system  with 
a  streptococcal  vaccine.  All  that  now  probably  remains  is  a 
rectal  fistula,  which  we  fail  to  close,  and  after  some  further 
extended  trial  we  give  vaccine  -  therapy  up  as  hopeless. 
We  may  even  scarify  the  sinus,  plug  and  dress,  and  yet  fail 
to  get  good  results.  We  make  a  microscopical  examination 
of  the  pus,  and  find  we  are  dealing  with  a  mixed  infection 
— for  example,  a  secondary  infection  by  staphylococci — 
and  in  addition,  being  near  the  anus  and  rectum,  perhaps 
B.  coli  have  become  established.  The  system  of  treatment 
has  been  condemned  in  failing  to  complete  a  cure,  but  how 
could  a  streptococcal  vaccine  cure  a  staphylococcal  and  coli 
infection  ?  In  short,  the  onus  of  failure  rests  with  the 
immunizer  in  this  case.  Again,  take  a  case  of  poll-evil, 
where,  the  infection  being  unusually  severe,  necrosis 
of  a  piece  of  deep-seated  tissue  follows,  which  would  in 


VACCINE-THERAPY  71 

the  ordinary  course  slough  out,  but  in  this  case  is  so 
imprisoned  that  separation  cannot  take  place.  Needless 
to  add,  a  vaccine  could  never  effect  a  cure  until  the  dead 
slough  had  been  removed,  and  vaccine-therapy  should  not 
be  condemned  because  it  failed  in  what  it  was  never 
intended  to  do — i.e.,  remove  a  dead  slough. 

Vaccines  are  divided  into  two  great  classes — stock 
vaccines  and  autogenous  vaccines.  A  stock  vaccine  is 
prepared  from  bacteria  of  the  same  species,  but  derived 
from  a  different  source,  and  kept  in  the  laboratory  for 
emergency  and  general  purposes.  An  autogenous  vaccine 
is  derived  from  cultures  of  the  actual  bacterium  or  its  strain, 
already  producing  the  infection  in  the  animal.  Again, 
vaccines  are  further  subdivided  into  monovalent  vaccines 
and  polyvalent  vaccines,  the  former  being  derived  from  a 
single  strain  of  a  particular  bacterium,  and  the  latter  from 
several  strains  and  races.  And,  lastly,  we  have  prophylactic 
and  curative  vaccines,  the  former  composed  of  devitalized 
bacteria,  which  are  injected  into  an  animal  whose  species 
are  susceptible  to  the  ravages  of  the  bacterium;  and  by  so 
doing  that  animal  becomes  partly  or  completely  immune 
to  the  disease  -  producing  influences  of  that  specific 
bacterium.  The  latter,  as  the  term  implies,  are  utilized  in 
rousing  the  bactericidal  elements  of  the  animal  body  to 
overcome  the  invading  organisms  and  their  death-producing 
influences.  It  is  only  reasonable  to  suppose  that  an  auto- 
genous vaccine  is  to  be  preferred  to  a  stock  one,  and  this 
is  borne  out  in  practice  with  few  exceptions;  when  one  is 
dealing  with  an  acute  disease,  such  as  pneumonia,  a  stock 
vaccine  should  be  administered  at  the  outset,  inasmuch  as 
it  takes  time  to  isolate  the  causative  organism  and  prepare 
an  autogenous  vaccine,  for  during  the  time  spent  in  prepara- 
tion the  patient  may  be  getting  rapidly  worse.  To  prepare 
an  autogenous  vaccine,  we  take  the  morbid  material  from 
the  patient,  with  careful  aseptic  precautions  to  prevent 
outside  contamination ;  and,  as  this  is  an  important  point,  it 
is  perhaps  as  well  to  record  briefly  the  method  of  so  doing 


72  CLINICAL  BACTERIOLOGY  AND 

in  cases  as  they  are  seen  in  practice.  If  we  desire,  there- 
fore, to  obtain  a  growth  from  a  nasal  discharge,  it  is  wise 
not  to  make  a  culture  from  the  pus  taken  from  the  anterior 
nares,  but  farther  back  in  the  nasal  cavity,  that  region 
being  more  certain  to  contain  the  specific  organisms  without 
so  much  contamination.  Take  a  piece  of  stout  wire  about 
15  inches  long,  and  fix  on  one  end  a  firm  piece  of  cotton- wool 
(sterile),  and  pass  into  the  nostril  (evading  the  false  nostril). 
Gently  rub  the  mucosa  with  the  pledget  and  withdraw 
it,  immediately  plunging  it  into  a  stout  sterile  test-tube. 
Push  the  free  end  of  the  wire  through  a  cork,  and  bring 
it  down  into  the  mouth  of  the  tube.  The  pledget  will  then 
be  firmly  fixed  in  the  test-tube.  If  it  is  the  urine  we 
desire  to  examine  for  bacteria,  the  fluid  should  be  with- 
drawn with  a  sterile  catheter  into  a  sterile  tube  and  corked. 
This  should  be  centrifuged,  the  supernatant  fluid  withdrawn 
by  a  sterile  pipette,  and  one  or  two  loopfuls  of  sediment 
placed  on  slides.  One- drop  of  blood  should  now  be  taken 
from  the  finger-tip  and  mixed  by  the  pipette  with  the 
urine  deposit,  and  spread,  dried,  and  stained.  The  blood 
supplies  the  albumin,  which  coagulates  on  heat  and  fixes 
the  bacteria  if  present.  In  cases  of  thoracic  effusion  or 
bursal  effusion  of  joints,  these  cavities  should  be  aspirated 
by  antiseptic  methods,  etc.,  and  the  fluid  placed  in  sterile 
vessels.  When  dealing  with  milk,  it  is  advisable  to 
asepticize  the  end  of  the  teat  with  tincture  of  iodine, 
draw  a  quantity  of  fluid  away  before  collecting,  after  which 
some  should  be  drawn  into  a  sterile  vessel,  care  being  taken 
not  to  touch  the  fluid  with  the  hand  or  fingers.  In  the  case 
of  abscesses,  etc.,  all  one  requires  to  do  is  to  smear  the  pus 
on  the  slide,  and,  if  too  thick,  distilled  water  should  be 
used  to  dilute  it.  It  is  good  practice  to  make  three  smears 
for  examination,  and  to  stain  them  with 

1.  Carbol-methylene  blue. 

2.  Gram's  method  (contrast  stain,  neutral  red). 

3.  Ziehl-Neelsen  method  (for  acid-fast  organisms). 


VACCINE-THERAPY  73 

Having  arrived  at  a  fair  conclusion  as  to  the  causative 
organisms,  we  proceed  to  cultivate  them.  An  agar  tube, 
a  blood-agar  tube,  gelatin  tubes  (two — slope  and  stab), 
and  one  or  two  anaerobic  tubes,  should  be  inoculated 
and  incubated.  Growth  may  be  detected  in  from  twelve 
to  twenty-four  hours.  But  if  any  colonies  fail  to  appear 
within  that  period,  incubation  should  be  continued  for 
at  least  three  days  before  the  tubes  may  safely  be  said 
to  be  sterile.  When  we  get  a  mixed  growth,  we  should 
start  to  grow  our  bacteria  on  Petri  dishes.  A  Roux 
bottle  is  also  useful  for  cultivating  bacteria.  A  first  pure 
subculture  is  best  for  the  preparation  of  a  vaccine.  A 
careful  microscopical  examination  should  now  be  made  of 
the  colonies  for  means  of  identification,  noting  purity,  etc. 
This  being  satisfactory,  5  c.c.  of  a  O'l  per  cent,  saline  (NaCl) 
solution  is  pipetted  into  the  tube  or  bottle,  and  the  whole 
gently  shaken.  If  some  of  the  colonies  are  difficult  to  dis- 
place, a  bent  sterile  platinum  wire  or  pipette  should  be  used 
to  rake  the  surface  of  the  medium,  taking  care,  of  course, 
not  to  graze  it.  We  now  have  a  bacterial  emulsion,  which 
should  be  poured  into  a  sterile  test-tube,  and  which  in  turn 
should  be  heated  about  the  middle  in  the  blow-flame,  and, 
when  sufficiently  melted,  drawn  out,  and  the  pointed  end 
sealed  by  the  flame  (as  described  on  page  50,  Fig.  23),  and 
placed,  point  upwards,  in  plasticine  or  in  a  tube-rack.  The 
emulsion  should  now  be  well  shaken  for  ten  minutes  to 
break  down  bundles  or  chains  of  bacteria.  To  assist  in  this 
process,  glass  beads  (sterile)  are  sometimes  inserted  also. 

Standardization. 

Having  obtained  our  emulsion,  we  now  proceed  to 
standardize  it,  and  for  this  purpose  several  methods  are 
adopted.  The  writer  has  followed  Wright's  method,  which 
is  as  follows  :  A  capillary  pipette,  6  inches  long,  with  a 
rubber  teat  fixed  to  the  open  large  extremity,  is  taken,  and 
marked  \  inch  from  the  end  of  the  capillary  extremity 
with  a  grease  pencil.     This  will  indicate  one  unit  volume. 


74  CLINICAL  BACTERIOLOGY  AND 

The  emulsion-tube  should  now  be  scratched  at  its  narrow 
end  with  the  glass-knife  and  broken  off,  but  before  doing 
so  it  is  well  to  sterilize  the  end  in  the  flame.  A  ligature 
should  now  be  placed  round  one's  finger,  and  the  finger 
pricked;  with  the  pipette  one  unit  volume  of  blood 
should  be  taken  up.  Then  a  column  of  air,  one  volume 
of  emulsion,  another  column  of  air,  and  one  volume 
of  diluent  (i.e.,  a  \  per  cent,  citrate  of  soda  in  a 
normal  saline  solution).  If  the  emulsion  is  very  thick, 
several  volumes  of  diluent  may  be  taken  up.  This  latter 
prevents  the  blood  coagulating.  The  teat  of  the  pipette 
should  now  be  pressed,  and  the  contents  expelled  on  to  a 
clean  slide,  taken  up  again  and  expelled  until  a  thorough 
mixing  has  taken  place.  As  soon  as  the  test-tube  containing 
the  emulsion  is  finished  with,  it  should  be  sealed  in  the  flame 
and  set  aside  for  sterilization.  Three  slides  should  now  be 
laid  in  front  of  the  operator;  these  slides  should  have 
their  surfaces  roughened  by  rubbing  with  emery-paper 
(Hubert's  00),  and  a  drop  of  the  mixture  placed  on  a  corner 
of  each  slide,  on  their  convex  surface  spread  out  with  the 
spreader,  and  allowed  to  air  dry.  They  are  then  placed  in 
a  trough  to  fix,  containing  a  saturated  solution  of  corrosive 
sublimate;  washed  thoroughly  under  the  tap,  and  stained 
with  filtered  carbol-thionin  for  two  minutes ;  washed  again, 
and  dried  with  filter-paper.  The  specimen  is  now  ready 
for  examination,  and  should  show,  if  satisfactory,  a 
fairly  even  field  of  bacteria,  with  blood -cells.  More- 
over, the  bacteria  should  not  be  in  bundles  or  chains. 
If  they  are,  the  emulsion  must  be  well  shaken  again;  or 
if  there  is  a  great  dearth  of  bacteria,  we  must  make 
a  stronger  emulsion,  or,  rather,  add  a  smaller  volume  of 
diluent,  and  go  over  the  whole  process  as  before,  and  make 
fresh  specimens.  Should  we,  however,  have  a  satisfactory 
field  (the  beginner  may  have  many  unsatisfactory  fields 
before  he  gets  a  satisfactory  one),  we  proceed  to  count  the 
bacteria  and  the  blood-cells  under  a  -^  oil-immersion  lens 
and  the  highest  available  eyepieces,  so  as  to  limit  the  field 


PLATE  VI. 


STANDARDIZATION    nF   BACTERIAL    EMULSION,    SHOWINI 
BACTERIA    AND    RED    BLOOD-CORPUSI   I  ES. 


To  face  [iwjr  74. 


VACCINE-THERAPY  75 

view  as  much  as  possible.  That  the  field  may  be  divided 
into  squares  to  facilitate  counting,  we  take  a  cover-glass  and 
mark  cross-lines  on  it  with  a  pen  and  ink,  or  four  hairs 
crossed  and  stuck  on  it  with  seccotine  will  answer  our 
purpose,  and  dropped  on  to  the  diaphragm  of  the  eyepiece. 
We  now  take  a  pencil,  and  count  in  each  square  the 
number  of  bacteria  in  one  column,  and  in  the  other  the 
number  of  red  corpuscles.  The  stage  is  moved,  and 
another  square  counted,  and  so  on,  until  we  get  a  total  of 
500  red  cells. 

Sometimes  we  find  a  square  contains  an  excess  of  bundles 
or  chains,  or  the  elements  may  be  badly  stained  and  set 
out.  It  is  then  better  to  pass  on  to  another  square,  and 
estimate  from  that.  The  two  columns  now  total  500  red 
corpuscles  as  against,  for  example,  640  bacteria ;  the  ratio 
is  therefore  500  to  640.  We  know  that  there  are  5,000,000 
red  cells  in  a  cubic  millimetre  of  blood.  Since  equal 
volumes  of  blood  and  emulsion  have  been  taken,  1  cubic 
millimetre  of  emulsion  will  contain — 

5,000,000x640  =  6  ^0,000. 
500  '      ' 

But  1  c.c.  contains  1,000  cubic  millimetres;  therefore  the 
emulsion  contains  6,400,000  x  1,000  =  6,400,000,000  bac- 
teria per  cubic  centimetre,  and  by  a  process  of  dilution  any 
suitable  strength  can  be  obtained.  Now  we  desire  to  make 
our  vaccine  for  future  use.  The  bottles  used  for  storing- 
purposes  are  rubber-capped,  25  and  50  c.c.  capacity, 
sterilized  and  filled  with  £  per  cent,  carbolic  and  NaCl 
(0*85  per  cent.)  solution;  and  we  wish  every  cubic  centi- 
metre to  contain  1,000,000,000  devitalized  bacteria,  there- 
fore in  our  50  c.c.  we  must  put  50,000,000,000  bacteria,  and 
as  we  have  already  seen  every  cubic  centimetre  of  our 
emulsion  contains  6,400,000,000  bacteria,  it  is  simply  a 
question  of  dividing  the  greater  amount  by  the  lesser  : 

50,000  million 


6,400  million 


=  7-81   (about). 


76  CLINICAL  BACTERIOLOGY 

We  therefore  must  extract  from  the  stock  vaccine  bottle 
diluent  solution  to  the  quantity  of  7*81  c.c,  to  make  room 
for  the  like  quantity  of  bacterial  emulsion,  which  has  been 
previously  drawn  up  by  the  syringe  to  be  injected  later  into 
the  stock  vaccine  bottle  by  passing  the  needle  through  the 
rubber  cap,  giving  us  a  standardized  dose  of  vaccine  of 
1,000,000,000  bacteria  in  every  cubic  centimetre  of  solution. 
Having  now  obtained  this  strength,  we  can  give  what 
quantity  we  desire — i.e.,  a  ^  c.c.  containing  500  million ; 
in  1  c.c,  1,000  million;  and  so  on. 

As  soon  as  we  have  made  an  emulsion  and  obtained  the 
exact  ratio  of  bacteria,  we  place  the  tube  in  a  moist  steri- 
lizer, to  kill  all  bacteria,  at  a  temperature  of  58°  to  60°  C. 
for  forty-five  minutes.  Care  must  be  taken  not  to  overcook, 
as  the  therapeutic  value  of  the  vaccine  is  reduced  thereby. 
On  the  contrary,  it  is  essential  to  kill  all  bacteria,  and  to  prove 
this  a  small  quantity  should  be  pipetted  into  a  culture-tube 
and  incubated.  If  no  growth  takes  place  in  from  twenty- 
four  to  forty-eight  hours,  one  may  take  it  the  emulsion 
is  sterile.  The  stock  bottle  into  which  the  vaccine  is  to  be 
placed  is  filled  with  (after  sterilization)  a  £  per  cent. 
carbolic  acid  and  0*85  per  cent,  salt  solution,  and  sterile 
rubber  caps  applied,  and  as  already  explained,  an  exact 
quantity  of  this  solution  has  to  be  withdrawn  from  the 
bottle  by  a  syringe,  the  needle  of  which  is  passed  through 
the  rubber  caps,  to  make  room  for  the  same  quantity  of 
emulsion,  which  is  also  injected  through  the  cap  with  the 
syringe.  To  make  sure  of  absolute  sterility,  the  needle 
should  be  passed  through  a  drop  of  lysol  put  on  the  cap 
each  time  it  is  punctured. 


CHAPTER  IX 

THE   SYRINGE 

Syringes  of  various  capacity  and  make  are  in  use.  Prob- 
ably a  1  c.c.  and  a  10  c.c.  capacity  syringe  are  the  two  most 
useful  sizes,  as  they  can  be  used  as  a  vaccine  and  serum 
syringe  respectively. 

They  should  have  metallic  or  glass  pistons,  which  are 
much  to  be  preferred  to  leather-washer  ones.  It  is  not 
necessary  to  have  a  large -bore  needle,  as  the  serums  and 
vaccines  are  very  fluid.  For  emergency  an  ordinary  hypo- 
dermic syringe  will  answer  one's  purpose. 

It  is  advisable  to  have  the  syringe  made  sterile,  although 
the  writer  must  confess  in  practice  to  having  made  several 
thousand  injections,  and  in  many  cases  failed  to  sterilize 
the  instrument,  and  does  not  recollect  having  ever  had  an 
abscess  at  the  seat  of  injection  or  any  other  sequelae,  but 
always  makes  it  a  point,  however,  to  cleanse  the  syringe 
after  use  by  aspirating  several  capacities  of  weak  lysol 
solution. 

Where  one  has  to  be  specially  particular,  and  in  practice 
one  really  cannot  be  too  particular,  the  syringe  can  be  con- 
veniently sterilized  with  hot  oil  in  the  following  manner: 
Take  a  tablespoonful  of  salad-oil  and  place  in  a  soup-ladle 
(the  latter  can  be  fixed  upon  a  stand),  float  in  the  oil  a  crumb 
of  white  bread.  Heat  over  a  Bunsen  burner  or  spirit  lamp 
until  bubbles  of  steam  begin  to  rise  from  the  crumb.  This 
will  indicate  a  temperature  equivalent  to  boiling  water. 
Now  take  up  a  syringeful  of  oil,  and  then  expel  it  into  the 
ladle  again;  continue  to  apply  the  heat  until  the  crumb 
77 


78 


CLINICAL  BACTERIOLOGY  AND 


becomes  brown.  A  temperature  of  about  140°  C.  has  now 
been  reached,  and  at  this  stage  the  oil  should  be  aspirated 
and  expelled  several  times,  when  the  syringe  will  be  found 
sterile. 

Seat  of  Inoculation. 

In  the  horse  and  ox  the  region  chosen  for  inserting  one's 
vaccine  or  serum  is  usually  one  easy  to  get  at,  and  where 


Fig.  32. — Vaccine  Syeinge. 

the  subcutaneous  tissue  is  plentiful.  Over  the  scapula  or 
midway  up  the  cervical  region  fulfils  both  qualifications. 
In  the  dog  the  most  convenient  situation  is  on  the  inside 
of  the  thigh,  for  here  the  skin  is  thin  and  nearly  devoid  of 
hair.     Where  many  injections  require  to  be  carried   out, 


Fig.  33.— Serum  Syringe. 

it  is  advisable  to  operate  on  one  side  and  then  on  the 
other,  and  upon  no  account  should  one  injection  follow 
another  in  exactly  the  same  spot. 

The  hair  should  be  clipped  at  the  seat  of  inoculation,  and 
the  skin  painted  with  strong  tincture  of  iodine  or  a  little 
pure  lysol  lightly  applied.    This  precaution  we  never  fail  to 


VACCINE-THERAPY  79 

carry  through.  The  syringe  is  now  charged  with  the  neces- 
sary dose  of  vaccine ;  it  is  usual  to  apply  a  twitch  in  the 
horse,  although  some  of  them  do  not  resent,  especially  if 
the  needle  is  a  small  one.  A  fold  of  skin  is  then  taken  up 
between  the  left-hand  finger  and  thumb,  and  the  needle 
firmly  plunged  through  the  skin.  It  is  advisable,  although 
not  so  essential  as  it  is  in  the  human  subject,  to  see  all  the 
air  is  expelled  from  the  fluid-free  end  of  the  syringe. 

At  the  seat  of  injection  there  may  or  may  not  be  a  little 
swelling,  and  in  the  case  of  the  dog  stiffness,  but  if  there  is 
it  usually  passes  off  in  a  day  or  two. 


CHAPTER  X 

PHENOMENA  FOLLOWING  ACTIVE 
IMMUNIZATION  BY  VACCINES 

The  immediate  result  consequent  upon  a  suitable  dose  of 
vaccine  injected  into  an  animal  is  a  fall  in  the  amount  of 
opsonin  present  in  the  serum,  apparently  due  to  the 
gathering  up  of  most  of  the  available  opsonin  to  the  bodies 
of  the  bacteria  introduced.  This  fall  in  the  index  is  called 
the  "negative  phase,"  and  occupies  a  period  of  from  twelve 
to  thirty-six  hours  usually ;  if  the  dose  is  very  large  it  may 
last  for  days,  if  very  small  it  may  be  eliminated  altogether. 
With  this  fall  of  the  index  there  is  usually  a  rise  in  tempera- 
ture ;  the  pulse  may  be  slightly  quickened,  and  there  may  be 
a  slight  constitutional  disturbance,  but  in  the  majority  of 
cases  there  are  no  appreciable  disturbances.  When  one  is 
dealing  with  a  local  lesion,  such  as  quittor,  poll-evil,  fistula, 
etc.,  an  increase  in  the  quantity  of  the  discharge  will  be 
noticed,  the  parts  become  more  swollen,  and  the  animal 
evinces  symptoms  of  increased  pain  locally.  These  phe- 
nomena usually  last  not  longer  than  twenty-four  hours,  and 
are  replaced  by  a  return  to  a  more  normal  condition ;  the 
pulse-beat  becomes  less,  also  the  temperature,  the  discharge 
lessens,  and  the  animal  is  brighter.  We  have  now  entered 
upon  the  positive  phase.  This  is  due  to  the  stimulus  given 
by  the  vaccine ;  fresh  supplies  of  opsonin  are  elaborated, 
during  which  the  opsonic  index  rises,  usually  taking  a  day 
or  two,  when  the  maximum  is  reached,  and  a  condition  is 
now  established  which  may  remain  for  a  day  or  two,  and 
is  called  the  "  phase  of  increased  resistance." 
80 


VACCINE-THERAPY 


81 


In  some  patients  this  phase  is  a  prolonged  one ;  in  others 
it  is  of  shorter  duration.  What  the  immunizer  should 
really  aim  at  is  to  have  the  positive  phase  as  long  as  possible, 
and,  of  course,  the  negative  phase  very  short  in  duration ; 
this  can  largely  be  attained  by  using  a  suitable  vaccine 
and  of  proper  dosage. 

In  very  acute  affections,  such  as  pneumonia,  one  cannot 
have  the  negative  phase  too  short,  in  which  case  it  is 
advisable  to  give  a  minimum  dose  at  the  outset ;  this  will 
give  a  quick  positive  phase,  and  another  larger  dose  should 
then  be  given  before  the  transient  positive  phase  passes  off. 


Increased 

resistance 

Fig.   34. — Chart   showing   the   Effects   of 
Vaccine. 


Suitable   Dose   of 


In  some  cases  the  period  is  not  more  than  twelve  to  eighteen 
hours. 

It  is  obvious,  in  dealing  with  an  acute,  depressing  disease, 
the  vaccine  will  produce  a  very  pronounced  negative  phase, 
and  the  patient's  resources  between  the  two  may  be  so 
exhausted  that  dissolution  will  follow.  This  is  naturally  a 
condition  the  immunizer  wishes  to  ward  against. 

It  is  therefore  a  wise  axiom  to  follow  :  Never  give  a 
maximum  dose  of  vaccine  in  an  acute  disease  with  rapid  pulse 
and  high  temperature,  remembering,  if  the  temperature 
is  high,  a  full  dose  of  vaccine  will  make  it  go  still  higher. 

The  tracing  given  in  Fig.  34  shows  what  really  should  take 
place  after  a  suitable  dose  of  vaccine  has  been  administered : 


82  CLINICAL  BACTERIOLOGY  AND 

When  the  dose  has  been  too  small,  we  get  an  immediate 
positive  phase  of  short  duration  and  of  little  or  no  curative 
value.     Thus : 


Fig.  85. — Dose  of  Vaccine  is  too  Small. 

On  the  contrary,  if  the  dose  has  been  too  large,  we  get 
an  immediate  fall  of  the  index.  Nature's  balance  has  been 
so  upset  she  cannot  assimilate,  so  to  speak,  the  dose;  great 
depression  sets  in,  and  several  days  may  pass  before  a 
restoration  to  the  previous  state  takes  place. 


Fig.  36. — Dose  of  Vaccine  is  too  Large. 

In  the  horse  we  have  never  noticed  this  phase,  and  have 
given  doses  very  many  times  greater  than  orthodox  present- 
day  teaching  suggests.  In  the  case  of  the  dog,  we  confess 
to  having  a  few  isolated  instances  of  apparent  temporary 
collapse,  etc.,  following  upon  vaccine-therapy,  but  never 
with  fatal  results. 

The  immunizer  should  aim  at  index  No.  1  as  his  ideal  to 
follow  out  in  practice. 

If  he  get  a  reaction  such  as  is  shown  in  index  No.  2, 
with  no  curative  results,  he  should  not  condemn  the  treat- 
ment, but  persevere  and  increase  his  dose. 

Index  No.  3,  in  the  hands  of  a  careful  practitioner, 
should  never  occur ;  and  if  it  does,  it  ought  to  teach  him  to 
be  less  heroic  and  more  careful  in  his  future  dealings  with 
vaccines. 

As  already  stated,  and  we  repeat  it  again  to  emphasize 
the  fact,  it  is  a  "sine  qua  non"  when  the  opsonic  index  falls, 


VACCINE-THERAPY  83 

the  temperature  invariably  rises ;  and  where  one  is  dealing 
with  an  acute  disease  and  a  high  temperature,  it  is  wise  to 
begin  with  a  minimum  dose  of  vaccine. 

When  the  temperature  is  very  high,  and  we  wish  to  begin 
the  vaccine  treatment,  we  give  a  big  dose  of  salicylic  acid, 
wait  three  to  five  hours,  and  then  inject.  This  we  find 
good  practice. 

Moreover,  it  is  advisable  not  to  give  a  second  dose  of 
vaccine  until  the  improvement  begins  to  fail.  In  subacute 
and  chronic  cases  this  varies  from  five  to  ten  days.  The 
second  and  succeeding  doses  should,  as  a  rule,  be  pro- 
portionately larger  than  the  first.  If  one  gets  no  response 
from  the  first  dose,  a  second  and  larger  dose  can  be  given 
almost  straight  away,  and  careful  notes  of  the  symptoms 
made. 

The  Dose  of  Vaccine. 

In  deciding  upon  the  most  suitable  dose  of  vaccine  for  a 
patient,  one  must  be  guided  by  circumstances. 

If  the  disease  is  an  acute  one  and  the  temperature  high, 
for  reasons  already  explained,  a  minimum  dose  must  be 
administered  at  the  outset;  on  the  contrary,  if  we  are 
dealing  with  a  chronic  condition  a  full  dose  is  indicated, 
and  is  quite  safe.  Again,  certain  bacterial  vaccines  are 
more  potent  in  their  actions  than  others.  For  example,  a 
given  streptococcal  vaccine  stimulates  the  bacteriolytic 
forces  much  more  effectively  than  a  staphylococcal  vaccine 
<>f  the  same  strength  does. 

In  judging  as  to  whether  a  suitable  dose  has  been  given  or 
not,  one  must  take  note  of  every  little  clinical  detail;  but  in 
reading  these  off  one  must  not  rely  upon  a  single  condition 
alone.  It  is  much  safer  to  total  them  all  together,  as  it 
were,  and  strike  averages.  For  example  take  a  case  of 
acute  pneumonia:  the  temperature  after  an  injection  may 
rise,  and  at  first  consideration  one  might  suspect  a  pro- 
nounced negative  phase  following  upon  the  vaccine  ;  but 
this  rise  may  largely  be  a  coincidence,  the  primary  cause 


84  CLINICAL  BACTERIOLOGY  AND 

being  probably  an  auto -intoxication  proceeding  from 
the  seat  of  infection. 

Again,  the  temperature  may  be  undisturbed,  while  the 
pulse  is  increased,  the  breathing  more  laboured,  and  pul- 
monary effusion  increased,  and  this  may  be  the  negative 
phase  of  the  cycle. 

Where  we  are  dealing  with  a  local  chronic  condition,  the 
clinician  is  probably  in  a  better  position  to  judge  the  actual 
state  of  affairs,  for  here  we  have  local  manifestations  upon 
which  to  make  our  observations  and  draw  our  conclusions. 
These  will  be  given  in  the  succeeding  chapters,  as  their 
respective  diseases  are  being  dealt  with. 

Although  one  has  to  use  a  certain  amount  of  discretion 
in  deciding  upon  dosage,  failure  to  effect  a  recovery  may 
solely  depend  upon  the  immunizer  being  too  careful  and 
using  too  small  a  dose.  On  the  other  hand,  we  may  say  the 
horse  and  ox  in  particular  are  very  tolerant  to  big  doses,  and 
make  excellent  subjects  for  vaccine-therapy.  These  con- 
clusions have  been  formed  after  watching  the  effects  of 
several  thousand  injections. 

On  the  dog  one  has  to  exercise  perhaps  a  little  more 
caution. 

Many  of  the  vaccines  advertised  at  the  present  day  are 
of  little  value,  if  only  for  the  fact  that  the  doses  are  too 
small;  we  have  had  repeatedly  to  give  half  a  dozen  in  one 
before  getting  a  suspicion  of  a  negative  phase. 

To  a  young  colt  seven  days  old,  suffering  from  a 
streptococcal  infection,  we  have  started  by  giving  as  a  dose 
100,000,000  devitalized  bacteria,  increasing  each  dose  until 
the  maximum  of  500,000,000  was  reached. 

If  a  young  colt  can  tolerate  these  doses,  how  much  more 
so  an  adult  horse  ! 

The  Age  of  Cultures. 

Because  of  involution  forms,  one  must  have  young 
cultures,  and  endotoxins  appear  to  be  best  suited  for 
vaccine-therapy  purposes. 


VACCINE-THERAPY  85 

The  following  is  a  table  of  the  average  culture  age  of 
some  of  the  more  common  bacteria  : 


Staphylococci' 

citreus  \ 
albus     L     ... 

...     12  to  48  hours. 

^aureus  ) 

Streptococci  

...     48  to  72  hours. 

Pneurnococci             

...     36  to  48  hours 

B.  coli  communis 

...     4  to  8  hours. 

Micrococcus  catarrhalis     ... 

...     5  to  10  hours. 

Micrococcus  letragenus 

...     5  to  10  hours. 

Bacillus  pyocyaneus 

...     24  to  36  hours. 

Dose  of  Autogenous  Vaccine  to  Horses  and  Cattle. 

The  horse  and  the  ox  are  very  tolerant  to  big  doses  of 
vaccines,  and  in  our  experience  make  excellent  subjects 
for  sero-vaccine  therapy. 

As  already  stated,  when  the  disease  is  running  a  very 
acute  course,  smaller  doses  as  a  rule  should  be  given ;  while, 
on  the  contrary,  in  an  animal  subject  to  a  chronic  bacterial 
invasion  the  system  is  more  tolerant  to  much  larger  doses. 

When,  however,  an  animal  is  suffering  from  an  acute 
staphylococcal  infection,  a  larger  proportionate  dose  of 
vaccine  can  be  given  here  than  can  be  given  in  the  case 
of  a  streptococcal  invasion  with  a  streptococcal  vaccine. 
After  three  years'  experience  of  immunization  with  auto- 
genous vaccines,  we  find  the  following  range  of  doses  to 
give  the  best  results,  but  we  would  point  out  the  immunizer 
must  ascertain  for  himself  to  what  extent  he  may  go  in 
dosage  with  each  individual  case,  remembering  that  not  only 
does  he  require  to  consider  the  idiosyncrasies  of  the  patient 
itself,  but  also  the  vaccine  he  is  using.  In  short,  there  is  no 
absolute  rule  of  thumb  to  go  upon. 

Initial  Dose. 
Minimum.  Optimum.  Maximum. 

Staphylococci  (three  kinds)...  250,000,000  500,000,000  750,000,000 

Streptococci 100,000,000  350,000.000  500,000,000 

Pneurnococci 75,000,000  250,000,000  500,000,000 

Bacillus  coli  grow? 200,000,000  350,000,000  750,000,000 

Micrococcus  catarrhalis      ...  250,000,000  350,000,000  500,000,000 

Micrococcus  tetragenus       ...  250,000,000  350,000,000  500,000,000 

Bacillus  pijocyanexis            ...  250,000,000  350,000,000  500,000,000 


CLINICAL  BACTERIOLOGY 


Dose  of  Autogenous  Vaccine  for  Adult  Dog  (Medium- 
Sized). 


Initial  Dose. 

Minimum. 

Optimum. 

Maximum. 

Staphylococci  (three  kinds) ... 

50,000,000 

125,000,000 

200,000,000 

Streptococci    

25,000,000 

100,000,000 

150,000,000 

Pneurnococci  ... 

20,000,000 

75,000,000 

150,000.000 

Bacillus  coli  group 

50,000,000 

100,000,000 

200,000,000 

Micrococcus  tetragenus 

50,000,000 

100,000,000 

200,000,000 

Bronchosepticus  (isolated  by 

Ferry)      

50,000,000 

100,000,000 

200,000,0U> 

CHAPTER  XI 

SERUMS  AND  THEIR  MODE  OF 
PREPARATION 

Seeo-therapy  is  a  branch  of  prophylactic  and  curative 
medicine  which  has  made  great  strides  within  recent  years, 
its  primary  object  being  the  neutralizing  of  liberated  toxins 
circulating  in  the  blood  and  lymph  streams.  These  toxins 
are  the  products  of  specific  bacteria  located  in  certain 
structures  of  the  animal  body,  and  the  serum  itself  does 
not  prevent  the  growth  of  the  bacteria. 

When  these  pathogenic  bacteria  settle  in  a  structure, 
they  incubate,  and  in  the  processes  of  growth  pour  out 
a  constant  stream  of  toxin,  which  is  continually  entering 
into  a  kind  of  combination  with  the  receptors  of  the  cells. 
It  therefore  follows  that  antidotal  serum  should  be  used 
before  serious  cell  changes  have  taken  place.  And  this 
is  exactly  borne  out  in  practice,  as,  for  example,  in  tetanus, 
swine  fever,  and  diphtheria  in  man ;  and  to  make  certain 
of  a  recovery  the  serum  should  be  injected  before  the 
diseases  are  established. 

As  the  making  of  serums  scarcely  comes  within  the 
domain  of  the  individual  practitioner,  a  general  outline  only 
of  the  modus  operandi  will  be  given. 

The  horse  is  the  animal  commonly  used  both  for  the 
making  of  human  and  veterinary  serums,  and  it  is  most 
essential  he  should  be  quite  healthy  and  be  free  from 
such  diseases  as  tuberculosis  and  glanders,  for  which  he 
is  tested  by  tuberculin  and  mallein  respectively. 
87 


88  CLINICAL  BACTERIOLOGY  AND 

The  animal  is  then  inoculated  with  a  dose  of  attenuated 
toxin  prepared  by  heating  the  virulent  poison  or  by  treat- 
ing it  with  a  chemical  agent,  either  of  which  will  reduce 
its  strength.  Repeated  inoculations  should  be  followed  up, 
at  stated  intervals,  with  increasingly  virulent  doses. 

The  toxins  which  are  injected  are  prepared  by  growing 
the  specific  bacteria  upon  suitable  fluid  media.  The  cultures 
are  then  passed  through  a  Pasteur-Chamberland  filter,  or  a 
similar  filter,  to  remove  the  bodies  of  the  bacteria. 

When  a  sufficient  degree  of  immunity  has  been  reached, 
and  three  or  four  days  have  elapsed  after  giving  the  last 
dose  of  toxin,  the  blood  is  withdrawn  under  aseptic  pre- 
cautions from  the  jugular  vein  by  a  sterile  cannula,  and 
collected  in  sterile  vessels.  These  are  put  aside,  preferably 
on  ice,  to  allow  the  blood  to  clot,  and  in  due  course  the 
serum  is  decanted  into  sterile  stock  bottles,  a  small  quantity 
of  preserving  {antiseptic  added,  and  they  are  ready  for 
use. 

Antibacterial  serums  are  produced  in  a  similar  manner, 
but  the  actual  bacteria  themselves  are  injected  instead  of 
their  toxins.  Before  a  degree  of  tolerance  is  reached,  the 
dead  bodies  of  the  bacteria  or  an  attenuated  culture  are 
given  at  the  outset,  or  a  dose  of  antitoxic  serum  is  ad- 
ministered to  mitigate  the  effects  of  the  first  virulent  dose. 

All  serums  should  be  tested  before  using,  to  see  that  they 
are  free  from  bacterial  contamination,  by  flooding  some  upon 
a  culture  medium  and  incubating,  and  to  insure  non- toxicity 
the  serum  should  be  injected  into  a  small  animal  first. 

Serums  are  standardized  by  the  physiological  test.  An 
animal  is  taken  which  reacts  in  a  constant  manner  to  the 
poison,  and  dying  within  a  certain  time.  It  is  known  that 
a  given  quantity  of  poison  will  destroy  an  animal  of  a  given 
weight  in  a  given  time.  This  is  taken  as  a  standard.  It 
is  then  necessary  to  find  what  amount  of  antitoxin  is  re- 
quired to  neutralize  this  dose,  and  we  find  equal  quantities 
of  a  given  antitoxin  will  do  this,  a  standard  thus  being 
set  up. 


VACCINE-THERAPY  89 

In  therapeutics  serums  may  be  used  alone  or  in  conjunc- 
tion with  vaccines — in  fact,  in  many  cases  the  combination 
is  a  happy  one,  particularly  in  the  diseases  attributed  to 
the  streptococcus.  Again,  in  some  diseases  we  are  com- 
pelled to  fall  back  upon  serums  alone,  as  there  are  certain 
specific  infections  where  the  causative  bacterium  has  never 
been  isolated :  hence  vaccines — at  least,  according  to  the 
present-day  meaning  of  the  word  "  vaccine  " — are  not  pro- 
curable. These  specific  diseases  belong  to  what  are  known 
as  the  ultravisible  virus  group,  of  which  the  following  are 
the  principal : 

Swine  fever. 

Canine  distemper  (probably). 

Rabies. 

Foot  and  mouth  disease. 

Specific  pleuro-pneumonia  (Bovine). 

Rinderpest. 

Variolas. 

Cape  horse  sickness. 

It  may  be  advisable  to  point  out  here  there  is  a  distinct 
phenomena  particularly  recognized  in  human  medicine 
following  upon  the  injection  of  sera  under  certain  cir- 
cumstances, and  it  is  known  as  "  serum  disease."  In 
animals  a  similar  condition  has  been  noted. 

Large  quantities  of  serum  may  be  injected  into  man 
almost  daily,  and  extending  to  two  or  three  weeks  without 
ill-effect.  If,  however,  a  period  of  twelve  or  more  days 
elapse  between  any  two  given  injections,  serious  results  are 
liable  to  follow.  The  condition  of  the  system  is  then 
hypersensitive  and  is  technically  know  as  anaphylaxis,  and 
in  some  cases  the  result  is  fatal  collapse. 

Theobald  Smith  noticed  when  guinea-pigs  were  injected 
with  horse  serum  and  a  second  injection  given  after  the 
elapse  of  ten  days,  the  pig  will  show  signs  of  hypersensi- 
tiveness,  and  if  the  dose  is  sufficient  death  may  follow. 

Again,  it  has  been  shown  that  the  sera  obtained  from  one 


90  CLINICAL  BACTERIOLOGY 

species  and  injected  into  another  species  of  animal  is  more 
likely  to  produce  anaphylaxis  than  if  the  sera  is  obtained 
from  the  same  breed  of  animal  we  desire  to  immunize. 
Thus  horse  serum  should  not  be  used  upon  the  ox,  more 
especially  if  a  considerable  lapse  of  time  is  allowed  to 
follow  any  two  injections,  and  vice  versa. 


CHAPTER  XII 

SPECIAL  DISEASES,  CAUSED  BY  SPECIFIC 
BACTERIA,  WHICH  ARE  SUITABLE  FOR 
TREATMENT  BY  SERO-VACCINE  THERAPY 

Diseases  affecting  the  Cutaneous  System. 

Disease.  Cause. 

Abscess        ...         ...         ...      Staphylococci,    streptococci,   B.    mallei, 

B.pyocyaneus,  B.  coli,  B.  tuberculosis, 
actinomycosis,  bothrioinycosis. 

Poll-evil  with  fistula        ...      Streptococci,  staphylococci,  B.  coli. 

Acne  ...         ...         ...      Acne  bacillus,  streptococci,  staphylococci. 

Mange  (Demodex  canis) ...      Staphylococci  (secondary  infection). 

Ulcers  B.    tuberculosis,   streptococci,   staphylo- 

cocci, pneuinococci,  B.  'pyocyaneus, 
B.  mallei. 

Abscess. 

In  practice  one  sometimes  finds  cutaneous  multiple 
abscesses  which  are  very  intractable  to  treatment  with  the 
ordinary  orthodox  therapeutical  dressings  and  drugs.  It  is 
in  cases  such  as  these  the  immunizer  can  do  much  for  his 
patient.  As  already  seen,  there  are  a  fair  variety  of 
bacteria  which  are  pus-producers  and  which  enter  into  the 
formation  of  abscesses.  Staphylococcus  albus  and  aureus 
are  the  two  bacteria  most  commonly  found  in  superficial 
abscesses ;  but  owing  to  the  situation  and  the  easy  means 
for  further  infection  one  often  finds  a  mixed  bacterial 
element. 

It  is  therefore  of  first  importance  the  immunizer  should 
become  conversant  with  the  nature  of  the  infection  at  the 
outset.  A  film  preparation  should  be  made  and  examined 
after  staining ;  a  specimen  should  also  be  stained  by  Gram's 


92  CLINICAL  BACTERIOLOGY  AND 

method,  and  the  results  compared.  Needless  to  add,  the 
pus  should  be  so  taken  that  there  is  no  danger  of  outside 
contamination  by  other  bacteria.  If  need  be,  a  stroke 
culture  on  agar  should  be  made,  and  a  gelatin-tube  also 
inoculated. 

Having  ascertained  the  causa  causans,  a  suitable  auto- 
genous vaccine  should  be  made  in  the  manner  already 
described. 

When  the  abscess  or  abscesses  are  very  tense  and  dis- 
tended with  purulent  matter,  internal  pressure  should  be 
relieved  by  lancing,  to  bring  about  lymphoid  osmosis.  Hot 
fomentations  should  be  applied  to  them  to  cause  capillary 
distension,  relieve  congestion,  and  increase  the  flow  of 
bacteriolytic  and  bacteriotropic  blood  to  the  parts ;  and  if 
the  blood  is  viscid,  citrate  of  soda  solution  should  be 
applied,  with  the  internal  administration  of  citric  acid. 
Upon  no  account  should  strong  antiseptics  be  applied, 
as  they  will  undo  all  the  good  the  vaccines  are  endeavour- 
ing to  do. 

If  the  abscesses  are  old-standing,  and  they  usually  are, 
one  can  begin  with  full  doses;  500,000,000  staphylo- 
cocci to  a  full-grown  horse  may  be  given  at  the  outset,  and 
the  results  carefully  watched.  In  five  to  seven  days 
1,000,000,000  may  be  given.  A  week  later  2,000,000,000 
may  safely  be  injected,  and  so  on,  increasing  the  dose  until 
one  finds  the  maximum  benefit  has  been  gained. 

In  the  case  of  streptococci,  one  cannot  give  such  large 
doses,  and  we  would  not  advise  giving  more  than  100,000,000 
as  an  initial  dose,  increasing  in  the  same  proportion  as  for 
staphylococci.  When  the  streptococcal  infection  is  a  severe 
and  acute  one,  we  have  obtained  considerable  benefit  by 
combining  the  vaccine  with  the  antistreptococcal  serum. 

Fistula   of  the   Poll   (Poll -Evil)   and   Fistula   of  the 
Withers  in  the  Horse. 

These  two  conditions  may  be  conveniently  taken  together. 
The  bacterial  cause   is  the  same,  the  pathological  lesions 


PLATE  VII. 


STAPHYLOCOI  CI. 
(Rose  and  Carless'  "Manual  of  Surgery.") 


BACILLUS    COLI    COMMUNIS.        X    1000. 
(Hewlett's  "  Bacteriology.") 


To  fact   pnar  P2 


VACCINE-THERAPY  93 

are  identical,  and  the  curative  treatment  is  the  same  in 
each  case. 

The  immunizer  may  be  called  in  to  see  a  case  at  a  stage 
when  there  is  a  considerable  accumulation  of  pus,  and  by 
reason  of  the  situation  the  purulent  matter  digests  the 
adjacent  healthy  structures,  burrowing  deeply — involving- 
vital  parts,  in  the  case  of  poll-evil,  the  spinal  cord  and  the 
spinal  vertebras — and  often  showing  no  disposition  to  burst. 
In  this  condition  one  finds  a  considerable  unilateral  or 
bilateral  swelling,  slightly  painful,  and  hot,  and  it  may  be 
fluctuating  or  tensely  hard.  In  such  a  condition  vaccine- 
therapy  would  be  of  little  or  of  no  use,  and  the  reason  is 
not  far  to  seek.  To  obtain  the  full  benefit,  the  antibodies 
of  the  blood  must  gain  access  to  the  focus  of  infection,  and 
where  there  is  a  considerable  internal  pressure  through 
excessive  purulent  accumulation  the  infiltration  of  lymph  is 
dammed  back.  The  blood  itself  may  be  very  rich  in  the 
necessary  bactericidal  elements,  and  yet  the  tissue  cells 
may  be  dying  from  nothing  short  of  bacterial  intoxication 
consequent  upon  auto-bactericidal  starvation. 

To  correct  this  condition,  the  pus  should  be  evacuated 
from  the  cavity  at  once,  when  the  curative  lymph  will 
flow  freely  through  the  diseased  area. 

If  this  fails,  we  must  look  for  other  causes.  The  lymph 
going  to  the  parts  may  coagulate  too  freely,  depositing 
itself  upon  the  cavity  wall  and  adjacent  tissues  as  a 
coagulum,  and  thus  acting  as  a  barrier  to  osmosis.  To 
remedy  this,  flush  the  cavity  with  a  solution  of  citrate  of 
soda,  and  give  internally  ounce  doses  every  five  hours  of 
citric  acid  until  a  free  flow  is  established. 

Again,  the  blood  itself  may  be  so  thick  and  tenacious  as 
to  undergo  too  rapid  and  easy  coagulation. 

Or  again,  if  the  case  is  one  of  long  standing,  the  abscess 
walls  may  be  so  thickened  and  indurated  by  the  deposit  of 
lymph,  etc.,  that  no  fresh  healing  lymph  can  pass  through. 
This  must  be  removed  by  the  free  use  of  the  curette. 

And,  lastly,  the  bactericidal  elements  of  the  blood  itself 


94  CLINICAL  BACTERIOLOGY  AND 

may  be  so  deficient  that  a  complete  regeneration  of  them 
is  required.  For  the  performance  of  this  function,  Ave  must 
look  to  vaccine  -  therapy  to  stimulate  the  deficiency  and 
supply  that  which  is  wanting. 

The  fact  of  an  animal  showing  symptoms  of  poll-evil  or 
fistula  to  the  skilled  practitioner  is  suggestive  of  one 
of  three  conditions  :  Either  the  patient's  opsonic  power  to 
the  causative  bacterium  is  too  low,  indicating  a  deficiency  of 
opsonins,  or  the  specific  infection  is  a  very  virulent  one,  or 
both. 

At  first  sight  it  seems  rather  strange  that  cutaneous 
bacterial  invasion  should  select  the  poll  and  the  withers  of 
horses  as  the  most  common  seats  of  entrance.  It  may  be 
there  are  three  distinct  forces  at  play  to  account  for  this  : 

1.  Some  localities  show  a  larger  percentage  of  cases  of 
fistula  and  poll-evil  than  others.  In  these  districts  one 
invariably  finds  bad  stables  and  low  roofs  and  unhealthy 
surroundings,  with  deficiency  of  sunlight. 

2.  Heredity  plays  an  important  part.  We  have  known 
three  generations  develop  poll-evil  and  fistula,  i.e.,  the  dam, 
the  daughter,  and  the  grandson,  the  latter  poll-evil  only. 
In  our  experience  the  opsonic  power  to  cutaneous  bacterial 
infections  is  markedly  low  in  some  families  compared  with 
others. 

3.  The  bacterium  most  commonly  found  in  purulent  poll- 
evil  and  fistulous  withers  is  the  Staphylococcus  albus,  in 
our  experience,  and  as  a  pathogenic  microbe  it  may 
be  defined  as  ubiquitous.  Certainly  the  horse's  skin, 
harness,  etc.,  harbours  this  organism  very  successfully. 
Anatomically  the  poll  and  the  withers  are  the  most  prominent; 
points  in  horses;  it  therefore  follows  with  low  roofs,  etc., 
bruising  is  more  liable  to  take  place  in  these  regions.  One 
sometimes  sees  in  practice  an  animal  receiving  a  blow  or  a 
bruise  in  these  parts;  an  inflammatory  swelling  takes 
place,  but  in  a  few  days  disappears.  This  the  writer  does 
not  call  poll-evil.  Clearly  here  the  skin  has  not  become 
broken  nor  bacterial  inoculation  taken  place;  but,  on  the 


VACCINE-THERAPY  95 

contrary,  should  bacteria  infect  the  bruised  area,  pus  soon 
forms,  and,  partly  by  gravitation  and  partly  by  digestion, 
burrows,  reaching  bursas  and  ligamentum  nuchas.  The 
ligament  and  the  bursas  being  lowly  vascular  organs, 
the  bactericidal  elements  going  to  these  parts  are  limited. 
This  gives  the  bacteria  a  good  chance  to  gain  a  footing, 
and,  when  once  established,  pus  increases,  burrows  down- 
wards, causing  local  death  to  many  tissue  cells,  and  even 
invading  the  osseous  structures  and  the  medullary  canal 
itself. 

The  Staphylococcus  aureus  may  be  found  in  conjunction 
with  the  albus,  and  sometimes  a  streptococcus  is  present. 
Very  rarely  have  we  found  a  Bacillus  coli.  The  latter's 
presence  is  more  commonly  discovered  in  fistula  of  the 
anus  or  deep-seated  wounds  in  the  region  of  the  rectum. 
To  make  an  autogenous  vaccine,  it  will  be  gathered,  the 
proper  bacteria  should  be  isolated  and  examined  and  cul- 
tivated, and  an  appropriate  vaccine  made. 

A  good  axiom  to  go  upon  in  practice  is  to  inject  one's 
vaccines  as  near  the  seat  of  disease,  if  it  be  a  local  one,  as 
possible.  And  we  even  go  farther  than  this  by  injecting 
the  vaccines  into  various  centres  round  the  swelling, 
if  circumscribed  ones,  such  as  poll-evil  or  fistula.  Opsonins 
are  largely  manufactured — it  is  believed,  in  muscular  and 
subcutaneous  tissue.  Allen  has  shown  that,  if  limbs  of 
animals  or  men  are  perfused  with  saline  solution  to  remove 
all  the  possible  blood-elements,  and  the  muscles  cooled  and 
minced,  and  their  plasma  extracted,  this  extract  has  a 
markedly  higher  index  than  that  of  blood-serum  towards  the 
various  bacteria — proving  at  least  muscular,  subcutaneous, 
and  intramuscular  tissue  contains  more  opsonins  than  the 
blood-serum  itself. 

The  writer  always  begins  by  injecting  500,000,000  staphy- 
lococci at  the  outset,  noting  the  symptoms,  and  when 
the  improvement  appears  to  be  drawing  to  a  close, 
follow  up  by  giving  1,000,000,000,  repeating  in  five  to 
seven  days  2,000,000,000. 


96  CLINICAL  BACTERIOLOGY  AND 

We  have  given  up  to,  as  a  dose,  10,000,000,000 
devitalized  bacteria,  after  a  decided  toleration  has  been 
established.  If  streptococci  are  present,  a  vaccine  from 
this  microbe  should  be  made,  and  the  initial  dose  should 
be  not  more  than  100,000,000.  One  has  to  be  more  careful 
with  a  vaccine  from  this  bacterium  than  with  staphy- 
lococci. In  five  to  seven  days  repeat  the  dose,  giving 
200,000,000.  As  a  maximum  dose,  after  several  adminis- 
trations, we  have  never  given  more  than  1,000,000,000, 
preferring  to  combine  the  vaccine  with  antistreptococcal 
serum,  if  the  case  requires  it. 

The  writer  has  had  one  troublesome  case  which  was  a 
mixed  infection  in  a  fistulous  withers  patient. 

The  wound  was  freed  of  staphylococci,  and  then  the  case 
came  to  a  standstill,  the  obstinate  bacterium  being  a  specially 
long-chained  streptococcus.  The  vaccine  failed  to  get  rid 
of  it  by  itself,  but  after  combining  it  with  streptococcal 
serum  the  patient  made  good  progress,  and  recovered. 

Clinical  Phenomena  after  Injection. 

In  these  cases  of  poll-evil  or  fistula,  one  has  the  advan- 
tage of  watching  local  conditions  which,  if  properly  inter- 
preted, are  of  singular  value  to  the  clinician. 

There  may  be  a  slight  constitutional  disturbance ;  this  is 
salutary.  Invariably,  if  the  vaccine  causes  a  response, 
the  local  swelling  increases  during  the  first  twenty-four 
hours ;  if  discharging,  the  discharge  also  increases,  contain- 
ing more  lymph  and  looking  more  healthy.  In  the  case  of 
poll-evil  the  head  is  carried  more  stiffly,  and  the  nose  more 
protruded. 

The  pus  should  be  carefully  noted.  The  writer  has 
found,  in  those  cases  where  the  opsonic  power  is  low  and 
the  bacterial  invasion  virulent,  the  purulent  elements  are 
of  a  greyish-yellow  colour,  thin  and  watery  in  consistence, 
and  possessing  little  or  no  viscosity.  The  clinician  may 
safely  interpret  this  condition  to  mean  the  lymphoid 
elements  of  the  blood  are  either  very  defective  in  them- 


VACCINE-THERAPY  97 

selves,  or  else  they  are  unable  to  reach  the  purulent  cavity. 
If  the  former,  suitable  vaccine  will  correct  the  defect ;  if 
the  latter,  surgical  aid,  as  already  indicated,  should  be  called 
into  action.  Invariably  these  cases  discharging  such  un- 
healthy pus  are  difficult  to  cure,  and  are  suggestive  of 
a  localized  necrosis ;  but  when  the  pus  becomes  thick  and 
tenacious,  or  what  our  forefathers,  who  made  few  errors 
in  the  field  of  practical  observation,  were  wont  to  call 
"  laudable  "  pus,  the  indications  are  most  favourable. 

The  aggravation  of  these  local  symptoms  after  twenty- 
four  hours  usually  decreases  when  the  positive  phase 
has  already  set  in,  and  from  this  period  an  improvement 
should  be  noted.  As  soon  as  this  appears  to  be  on  the 
wane,  another  and  larger  dose  should  be  given.  Needless 
to  add,  poll-evil  and  fistula  are  debilitating  diseases,  and 
good  food,  good  surroundings,  etc.,  are  essential  to  the 
making  of  a  good  recovery. 

A  word  of  warning  may  suitably  be  given  here.  The 
practitioner  may  examine  his  case  carefully,  make  an 
accurate  diagnosis,  prescribe  a  suitable  vaccine,  and  yet 
fail  to  bring  about  recovery. 

There  is  no  system  of  treatment  which  has  not  its 
limitations,  and  vaccine-therapy  is  no  exception  to  this 
rule.  The  immunizer,  with  industry,  skill,  and  adapta- 
bility, can  drive  these  limitations  farther  away,  giving 
himself  more  scope  to  work,  and  in  the  end  obtaining 
better  results.  But  there  are  other  points  to  consider. 
For  example,  one  practitioner  gives  up  a  case  after  a 
course  of  vaccine  treatment,  and  condemns  the  system 
in  toto ;  another  practitioner  would  probe  the  fistula, 
and  detect,  perhaps,  a  calcifying  necrosis  on  the  lower 
border  of  the  funicular  portion  of  the  ligamentum  nuchas, 
or,  maybe,  a  specific  periostitis  of  the  body  of  the  atlas  or 
axis.  Now,  the  most  perfect  autogenous  vaccine  and  the  most 
skilled  immunizer  would  prove  both  miserable  failures  if 
they  followed  up  vaccine-therapy  and  failed  to  curette  the 
necrosed  area  or,  maybe,  remove  a  dead  slough. 

7 


98  CLINICAL  BACTERIOLOGY  AND 

Again,  it  is  a  deep-rooted  idea,  and  even  with  men  of 
great  practical  experience,  to  go  on  irrigating  sinuses 
and  cavities  with  antiseptic  dressings  of  considerable 
strength,  their  logic  being  that  by  so  doing  they  are  killing- 
out  the  bacteria  present  in  the  cavity,  and  by  getting  rid 
of  the  cause,  they  reason,  the  effect — i.e.,  the  disease — will 
disappear.  It  requires  only  a  moment's  consideration  to 
see  that  such  reasoning  is  entirely  wrong ;  nay,  more,  the 
facts  are  not  borne  out  in  practice.  To  begin  with,  the 
most  vital  and  active  bacteria  are  not  to  be  found  in  the 
debris  of  an  abscess  or  in  the  centre  of  a  sinus.  Many 
of  them  are  degenerates,  and  therefore  possess  only 
slight  pathogenic  powers.  Some  are  devoured  by  the 
phagocytes.  Others  are  rendered  inert  through  the  action 
of  the  antibodies  and  the  poisonous  effects  of  their  own 
toxins.  But  in  the  walls  of  these  compartments  they 
are  most  active  and  virulent,  for  there  they  find  what  they 
want — heat,  moisture,  and  nourishment,  and  last,  but  not 
least,  removal  from  their  own  exotoxins.  Now,  how  far 
does  an  antiseptic  fluid,  such  as  an  aqueous  solution  of 
hyd.  perchlor.,  penetrate  into  the  walls  of  these  cavities, 
which,  be  it  remembered,  are  varnished  all  over  with  the 
exuded  lymph  ?  Moreover,  the  mercury  salt  frustrates  its 
own  ends  as  an  antiseptic  by  coagulating  the  albuminous 
lymph  and  making  it  a  still  more  perfect  varnish.  To  our 
mind,  there  is  nothing  which  detracts  more  from  the  healthy 
healing  of  a  wound,  and  hampers  Nature's  efforts  so 
completely,  as  the  repeated  use  of  strong  or  moderately 
strong  antiseptics.  If  an  antiseptic  has  to  be  used,  it  is 
much  better  practice  to  use  one  caustic  plugging  to  slough 
out  the  germ-laden  wall,  leaving  a  healthy  granulating 
surface  to  complete  the  repairing  process.  But  even  this 
process  has  its  drawbacks,  and  must  be  used  with  care.  The 
fact  of  applying  dressings  strong  enough  to  cause  a  slough 
not  only  causes  local  death  to  bring  about  the  slough,  but 
absorption  must  take  place  to  the  surrounding  healthy 
structures,  which  may  be  diffused  for  several  inches.     All 


VACCINE-THERAPY  99 

through  this  area  the  protective  bodies  and  tissue  cells  have 
had  their  vital  energies  depressed  by  the  poison,  and  their 
offensive  and  defensive  powers  consequently  lowered,  giving 
the  bacteria  a  chance  to  gain  another  and  extended  footing. 
The  only  antiseptic  solution  the  writer  uses  now  for 
these  cavities  is  weak  boric  lotion,  and  great  results  will 
follow  the  free  use  of  cold  tap-water  irrigations.  Normal 
saline  solution  also  makes  an  excellent  dressing  as  a  mild 
antiseptic,  tissue  stimulant,  and  restorative. 

Fistula  of  the  Presternum  of  the  Horse. 

This  condition,  in  our  experience,  is  not  very  common ; 
having  only  seen  six  cases,  and  three  of  these  have 
occurred  within  the  last  four  years.  The  condition  is 
caused  by  the  animal  usually  running  against  some  hard, 
fixed  object,  and  bruising  the  structures  from  the  skin  to 
the  cariniform  cartilage.  A  hard  or  semi-hard  swelling 
results ;  the  animal  walks  stiffly,  with  fore-legs  held  wide 
apart.  In  course  of  time — usually  a  considerable  time — 
an  abscess  forms,  points,  and  bursts,  the  discharge  being 
of  a  thin,  glairy,  sometimes  brown  or  dark-grey  colour. 
The  swelling  tends  to  subside  if  the  discharge  is  sufficient, 
and  the  animal  then  goes  better;  the  pus  may  cease  flowing, 
the  wound  closes,  and  the  swelling,  pain,  stiffness  return, 
and  when  a  sufficient  purulent  accumulation  has  taken  place 
the  abscess  again  points  and  bursts,  and  these  processes 
go  on  indefinitely.  It  is  probable  there  is  no  case  of 
spontaneous  cure  on  record. 

As  with  poll-evil  and  fistulous  withers,  we  have  here  also 
the  same  forces  at  work  and  the  same  anatomical  drawbacks 
which  Nature  encounters  in  her  endeavours  to  bring  about 
recovery — i.e.,  pus-producing  bacteria  on  the  one  hand,  and 
the  deficient  blood-supply  going  to  the  injured  parts  on 
the  other.  Sometimes  one  sees  a  solitary  sinus,  at  other 
times  there  may  be  two  or  three,  leading  either  to  one  or 
more  foci  in  the  sternum. 

As  with  all  other  bacterial  invasions,  so  here  one  must 


100  CLINICAL  BACTERIOLOGY  AND        » 

ascertain  what  causative  organisms  are  present  by  adopting 
the  usual  technique.  The  Staphylococcia  albus  and  aureus 
combined  we  have  found  in  our  three  last  cases.  We 
are  at  present  treating  a  case  where  we  detected  these  two 
bacteria,  and  in  addition  a  rather  uncommon  bacterium — 
i.e.,  B.  pyocyaneus  (belonging  to  the  chromogenic  class). 

In  this  disease,  one  has  to  give  full  vaccine  doses  if  the 
best  results  are  to  be  obtained.  The  writer  usually  starts 
with  750,000,000  Staphylococcus  aureus  and  albus,  and  in 
this  latter  case  combined  it  with  250,000,000  B.  pyocyaneus, 
the  doses  being  doubled  each  time. 

The  patient  has  had  up  to  the  present  three  injections, 
and  progress  so  far  is  quite  satisfactory :  the  swelling  is 
becoming  smaller  and  softer,  and  the  stiffness  has  dis- 
appeared. During  the  negative  phase  this  increased,  how- 
ever, and  for  twenty-four  hours  the  animal  scarcely  walked 
at  all,  being  very  stiff  on  his  front-legs.  Locally  there  is  one 
central  sinus,  which  discharges  intermittently.  The  sinus 
has  been  curetted  and  irrigated  with  soda  citrate  solution 
alternated  by  normal  saline  solution.  Since  these  notes 
were  made,  this  case  has  completely  recovered  and  been  in 
constant  work  for  six  months. 

Fistula  in  the  Region  of  the  Anus  and  Rectum. 

This  condition  in  equines  is  seen  often  as  a  sequel  to 
strangles  running  an  irregular  course,  and  is  most  intractable 
to  the  ordinary  forms  of  treatment.  If  the  case  is  one 
of  long  standing,  the  immunizer  usually  finds  the  purulent 
discharge  contains  more  than  one  bacterium.  Strepto- 
cocci there  usually  are,  staphylococci  there  may  be,  and, 
considering  the  situation,  one  has  every  justification  in 
being  on  the  careful  lookout  for  B.  coli  communis  ;  and  if 
present  a  suitable  vaccine  should  be  made.  In  human 
vaccine-therapy,  we  understand,  a  stock  vaccine  of  the 
B.  coli  acts  very  well. 

The  writer  recalls  treating  a  case  with  staphylococcal  and 
streptococcal  vaccine,  giving  extremely  large  doses  and  fail- 


VACCINE-THERAPY '  101 

ing  to  close  the  sinuses.  A  further  microscopical  and 
biological  examination  was  made,  and  we  found  a  few 
shrivelled  staphylococci,  but  an  abundant  display  of  very 
active  B.  colt. 

The  initial  doses  in  a  case  of  this  kind  should  be — Strepto- 
cocci, 100,000,000;  staphylococci,  500,000,000;  B.  coli, 
100,000,000  :  repeated  in  five  to  seven  days,  according  to 
conditions,  in  appropriately  increasing  doses.  Of  course, 
if  there  is  any  necrosed  area  in  the  sinus,  this  should 
be  removed,  as  Nature's  sloughing  efforts  in  these  cases 
are  very  indolent.  Here  also  oft  -  repeated  antiseptic 
dressings  should  be  deprecated. 

Acne — Pustular  Dermatitis  Contagiosa. 

This  condition  is  seldom  seen  in  equines  in  this  country. 
It  occurs  in  the  form  of  papules  or  pustules,  usually  on  the 
parts  of  the  body  where  the  harness  rubs.  It  is  due  to 
a  bacillus  which  sets  up  a  purulent  inflammation  of  the 
skin,  destroying  the  hair  follicles,  and  may  spread  all  over 
the  trunk  by  means  of  the  harness,  grooming  utensils,  and 
clothing.  The  presence  of  the  acne  bacillus  and  the  skin 
lesions  it  sets  up  seem  to  facilitate  the  growth  of  the 
ubiquitous  staphylococcus,  which  invariably  is  found  in  the 
purulent  discharges,  aggravating  the  disease  and  prolong- 
ing the  condition.  Streptococci  may  also  be  present  in  the 
pustules.  The  bacillus  is  anaerobic — at  least,  at  the  outset 
— and  is  Gram-positive. 

It  is  small  and  ovoid,  occurring  singly  or  in  short  chains. 
It  is  non-motile.  In  a  gelatin  stab  culture  small  colonies 
occur ;  on  glucose  agar  white  colonies  grow  very  slowly. 

In  this,  as  in  all  other  mixed  infections,  the  off  ending- 
bacteria  should  be  isolated,  and  a  vaccine  made  from  each. 

To  obtain  a  pure  culture  of  the  acne  bacillus,  take  a 
scraping  from  the  deeper  parts  of  the  sebaceous  plugs,  and 
mix  the  material  in  broth,  incubating  anaerobically  for  ten 
to  fourteen  days.  Should  staphylococci  be  present,  by  this 
time  they  will  be  inert,  while  the  acne  bacilli   will  have 


102  CLINICAL  BACTERIOLOGY  AND 

multiplied.  To  10  c.c.  of  melted  3  per  cent,  agar  rendered 
acid  by  hydrochloric  acid,  add  5  c.c.  of  fresh  sterile  ox 
serum,  and  pour  into  a  Petri  dish. 

Now  spread  a  loopful  of  the  acne-bacilli-laden  broth  over 
the  surface  of  the  plate,  and  incubate  for  three  days 
(anaerobically).  These  may  be  picked  off,  replated,  and 
incubated  aerobically,  and  a  bacterial  emulsion  made  in  the 
usual  way  preparatory  to  the  making  of  the  vaccine. 

Follicular  Mange  in  the  Dog. 

This  troublesome  skin  disease  of  the  dog  is  due  primarily 
to  the  burrowing  of  the  Demodex  folliculorum. 

In  course  of  time  the  protective  forces  of  the  skin  suffer 
considei-ably,  the  animal's  health  becomes  impaired,  his 
opsonic  index  falls,  and  the  patient  is  ripe  for  any  bacterial 
invasion  that  may  come  along. 

On  the  surface  of  the  skin  is  to  be  found  an  excellent 
medium  for  bacterial  growth — blood-serum  ;  and  at  a  body 
temperature  staphylococci  soon  manifest  their  presence  by 
excessive  pus  formation — so  much  so  that,  in  advanced 
cases,  the  writer  strongly  believes  one's  greatest  concern,  so 
far  as  a  cure  goes,  is  to  get  rid  of  the  bacteria,  and  that 
the  acari  are  but  secondary  factors  at  this  stage  in  the 
causation  and  continuation  of  the  disease. 

An  animal  in  such  a  condition  is  clearly  one  for  the 
immunizer,  provided  he  can  use  an  autogenous  vaccine. 
The  obstinate  nature  of  these  cases  toward  orthodox  thera- 
peutics lies,  not  so  much  in  the  fact  that  the  acari  and  the 
bacteria  are  so  difficult  to  destroy,  but  rather  they  are  so 
ungetatable  (if  one  may  use  the  word),  their  unassailable 
seat  being  principally  at  the  roots  of  the  hair  follicles.  It  is 
quite  obvious,  then,  to  reach  them  one  must  use  stro:ig 
penetrating  poisons,  which,  as  already  indicated  (in  previous 
chapters),  when  dealing  with  bacterial  infections,  is  to  be 
deprecated,  destroying  as  they  do  the  very  elements  we 
wish  to  preserve  and  fortify,  i.e.,  the  bactericidal  antibodies 
of  the  blood. 


VACCINE-THERAPY  103 

What,  then,  is  the  orthodox  therapeutist's  misfortune 
should  be  the  vaccine-therapist's  opportunity ;  for  the  fact 
that  the  bacteria  are  deep-seated  gives  him  a  better 
opportunity,  with  his  vaccine,  of  driving  home  in  full 
force  Nature's  whipped-up,  refreshed,  and  restored  anti- 
bodies. 

Although  staphylococci  are  the  usual  pus  -  forming 
bacteria  present,  and  particularly  the  albus,  it  does  not 
follow  other  bacteria  may  be  absent.  It  therefore  behoves 
the  practitioner  to  make  sure  by  careful  microscopical  and 
biological  examination  what  the  nature  of  the  infection  is. 

The  dose  of  staphylococci  depends  largely  upon  the  size, 
age,  and  general  condition,  of  the  dog.  To  a  medium- sized 
dog  we  usually  begin  with  75,000,000  devitalized 
bacteria,  watching  the  symptoms.  If  one  finds  a 
sufficient  negative  phase  has  been  established,  wait  about 
five  to  seven  days,  and  repeat  by  giving  100,000,000,  the 
third  dose  being  increased  to  150,000,000.  After  tolera- 
tion has  been  established,  we  have  given  to  a  retriever  as 
much  as  1,000,000,000  staphylococci.  If  the  improvement 
is  not  as  much  as  one  would  anticipate,  do  not  hesitate  to 
give  bolder  doses;  but  it  is  always  well  to  begin  with  a 
moderately  small  initial  dose,  and  work  upwards. 

Locally  one  must  endeavour  to  get  the  lymph  to  flow 
freely,  and  great  benefit  is  to  be  derived  from  boric  lint 
soaked  in  boiling  water,  applied  to  the  skin  very  hot 
and  over  this  a  thick  layer  of  oilskin  should  be  applied 
and  retained.  Citrate  of  soda  and  boric  lotion  should  also 
be  applied  at  intervals,  and  if  need  be  citric  acid  should  be 
administered  internally.  Later  a  course  of  calcium  sul- 
phide tabloids  should  be  carried  out.  Needless  to  add,  the 
patient  should  be  fed  up,  and  kept,  by  being  placed  in 
healthy  surroundings,  from  reinfecting  himself. 

The  opsonic  index  will  be  very  low  usually ;  in  one  case 
we  found  it  as  low  as  0"39.  This  was  a  chronic  case,  and 
the  animal  was  in  a  cachexous  condition. 


104  CLINICAL  BACTERIOLOGY 

Ulcers. 

Ulcer  formations  may  be  seen  in  any  part  of  the  integu- 
ment. 

They  may  be  single  or  multiple,  benign,  and  heal  with 
little  effort  on  the  part  of  Nature  or  the  practitioner.  They 
may  be  malignant,  such  as  cancerous,  glanderous,  tuber- 
culous, and  incurable. 

Needless  to  add,  vaccine -therapy  in  the  lower  animals 
excludes  these  three  latter  conditions. 

When  one  finds,  however,  a  simple  ulcer  becoming  con- 
taminated by  pathogenic  bacteria,  and  assuming  serious  pro- 
portions, with  probable  symptoms  of  septic  infection  or 
intoxication,  the  immunizer  can  do  much  to  relieve  suffer- 
ing and  save  life  with  suitable  vaccines. 

Invariably  the  infection  is  a  mixed  one,  and  this  point 
must  be  definitely  settled  before  curative  progress  can  be 
embarked  upon. 

Locally  the  curette  should  be  used,  and  even  a  single 
caustic  dressing  may  stimulate  healthy  granulation,  although 
it  must  not  be  repeated. 

When  the  deep  structures  of  the  corium  become  involved, 
and  if  the  infection  is  a  virulent  one,  localized  death  of 
tissue  may  take  place.  This,  of  course,  must  be  removed 
by  hot  fomentations,  the  lancet,  and  free  drainage. 

One  sometimes  meets  in  practice  a  destructive  and 
troublesome  ulcerative  keratitis  in  valuable  dogs  after 
distemper,  and  one  would  think  a  suitable  autogenous 
vaccine  would  be  of  great  service  here,  saving  eyes  and 
eyesight,  which  are  often  lost.  Of  this,  however,  the  writer 
has  no  experience,  and  only  throws  out  the  suggestion  should 
others  come  across  a  suitable  case  to  try  vaccine-therapy 
upon. 


CHAPTER  XIII 

BACTERIAL  DISEASES  AFFECTING 
SYNOVIAL  JOINTS 

Disease.  Cause. 

Traumatic  arthritis Staphylococci,  streptococci. 

Pyaernic  arthritis  in  foals     ...      Staphylococci,    streptococci     B.    coll, 

B.  pyocyaneus. 
Septicemic  arthritis  in  cattle      Streptococci. 

Traumatic  Arthritis. 

The  articular  elements  of  the  animal  body  become  the 
seat  of  bacterial  invasions  through  two  distinct  sources — 
i.e.,  first,  internally,  and  what  might  be  called  "auto- 
infection,"  the  bacteria  being  carried  usually  by  the  blood 
or  lymph  stream;  and,  secondly,  externally,  as  illustrated 
by  traumatism. 

Whatever  may  be  the  channel  by  which  bacteria  reach 
these  joint  cavities,  one  cannot  gainsay  the  fact  that  when 
they  do  get  there  they  find  an  admirable  breeding-ground, 
and  this  fact  is  soon  demonstrated  in  practice  by  the  very 
rapid  manner  in  which  they  develop,  disintegrating  the 
vital  structures  of  the  joint  and  adjacent  tissues. 

In  cases  of  traumatic  arthritis  the  invading  bacteria  usu- 
ally belong  to  the  staphylococci  group.  The  practitioner 
here  is  not  confronted  so  much  with  the  question  of  the  life 
and  death  of  his  patient  as  he  is  with  the  great  desire  of 
limiting  the  destructive  metabolic  changes  occurring  in  the 
joint  and  caused  by  these  pathogenic  bacteria.  When 
destruction  of  the  sensitive  articular  surface  takes  place, 
105 


106  CLINICAL  BACTERIOLOGY  AND 

anchylosis  is  certain  to  follow,  and  that  to  an  animal  of  the 
equine  species,  of  course,  means  destruction. 

We  therefore  maintain,  if  the  practitioner  is  called 
in  sufficiently  early — i.e.,  before  destructive  changes  have 
taken  place  within  the  joint — and  he  adopts  a  rational 
course  of  treatment  with  suitable  vaccines,  combined  with 
thermic  assistance,  he  need  fear  no  organic  stiffening  of  the 
joint.  Of  course,  where  pus  has  been  discharging  for  days, 
and  the  whole  of  the  articular  surface  has  been  stripped  by 
the  digesting  action  of  the  bacteria,  vaccine-therapy  will 
fail  to  restore  such  a  joint  to  a  normal  condition,  although, 
undoubtedly,  it  would  facilitate  the  removal  of  the  bacteria. 

Retardation  of  the  development  of  bacteria  within  the 
joint  is  effected  by  a  more  or  less  continuous  spray  of  cold 
water,  and  when  the  circulation  is  interfered  with,  or  in 
any  way  becomes  defective,  the  application  of  Bier's  treat- 
ment, by  driving  an  excess  of  bactericidal  elements  to  the 
joint,  forms  an  excellent  adjuvant  to  vaccine-therapy.  In 
addition  we  have  used  for  some  years  injections  of  ol. 
caryoph.,  with  excellent  results  in  open  joints,  and,  unlike 
most  antiseptics,  it  does  not  appear  to  irritate  or  upset  the 
local  tissues  it  comes  in  contact  with;  on  the  contrary,  it 
seems  rather  to  stimulate  than  depress  Nature's  restorative 
and  protective  elements. 

Pyaemic  Arthritis  as  seen  in  Young  Foals. 

The  etiology  of  the  condition  as  seen  in  practice  is  far 
from  being  complete.  Quite  a  number  of  bacteria  have 
been  isolated  as  causing  the  disease,  and  not  a  few  of  these 
are  undoubtedly  of  secondary  origin. 

It  appears  to  the  writer  we  have  here  two  distinct  con- 
ditions— i.e.,  bacterial  invasion  in  utero  and  umbilical  infec- 
tion after  birth. 

In  the  former  case  the  navel  cord  looks  macroscopieally 
healthy  and  shows  little  or  no  retrogressive  changes; 
nevertheless,  within  a  few  hours  after  birth  an  effusive 
arthritis  has  taken   place.     To  those  who  hold  infection 


VACCINE-THERAPY  107 

takes  place  from  the  navel  after  birtli  in  these  cases,  one 
must  reasonably  point  out  the  short  period  which  has 
elapsed  between  parturition  and  the  first  manifestation  of 
organic  disease  in  a  distant  joint.  Moreover,  in  practice  we 
have  ligatured  the  navel  cord  at  birth  with  sterile  catgut, 
painted  the  stump  with  tincture  of  iodine  and  then  with 
collodion,  and  repeated  the  dressing  daily,  keeping  the 
animal  under  the  most  perfect  hygienic  surroundings 
possible,  and  yet  within  a  varying  but  usually  short  period 
specific  arthritis  develops.  In  those  cases  the  animals  are 
nearly  always  weakly  at  birth,  and  if  they  live  they  seldom 
become  valuable,  being  stunted  in  their  growth,  etc. 
The  majority,  however,  die  from  what  appears  to  be  a 
pronounced  septicaemia. 

Where  joint-evil  occurs  through  bacterial  invasion  of  the 
navel  cord  after  birth,  one  usually  finds  an  extended  in- 
cubation period,  or  at  least  an  apparently  extended  period, 
for  one  cannot  state  definitely  when  the  navel  stump  may 
become  infected.  It  is  reasonable,  however,  to  presume 
the  older  the  animal  becomes,  the  more  the  cord  shrinks 
and  dries,  the  less  likely  one  is  to  get  bacterial  growth  and 
infection  from  this  source. 

Some  navel  cords  appear  to  be  more  predisposed  to  act 
as  bacterial  incubators  than  others ;  those  which  are  thick 
and  gelatinous  at  birth,  with  a  more  or  less  serous  exudate 
emanating  from  them  continually,  are  more  likely  to  lead  to 
joint-evil  than  small,  dried-up,  and  attenuated  cords. 

With  the  class  of  cases  which  appear  to  be  affected  in 
utero  we  must  confess  vaccine-therapy  in  our  hands  has 
been  of  little  or  no  service,  and  the  reason  for  this  seems 
to  be  we  have  here  a  bacterial  infection  primarily  affecting 
the  blood-stream — i.e.,  a  typical  septic  infection. 

Where,  however,  the  animal  appears  to  have  become 
contaminated  when  two  or  three  weeks  old,  provided  the 
bacteria  causing  the  condition  are  isolated,  and  an  autog- 
enous vaccine  made,  good  hopes,  if  the  treatment  is  started 
fairly  early,  of  a  cure  may  be  anticipated. 


108  CLINICAL  BACTERIOLOGY  AND 

Before  the  writer  gave  vaccine  -  therapy  the  serious 
consideration  he  now  does,  he  obtained  considerable  relief 
in  cases  where  the  joint  or  joints  were  greatly  distended 
with  turbulent-looking  synovia,  etc.,  by  aspirating  them, 
and  giving  the  fluid  to  the  patient  per  oram,  believing  in  this 
manner  the  patient  elaborated  its  own  protective  vaccine. 
Certainly  in  some  cases  the  animal  often  made  a  good 
recovery,  and,  to  say  the  least,  aspirating  the  joint  relieved 
internal  pressure,  thereby  reducing  the  painful  condition  of 
the  joint,  which  in  such  young  animals  so  often  kills  by 
exhaustion  alone.  Moreover,  we  found,  if  the  joint  was 
aspirated  moderately  early  in  the  course  of  the  disease,  one 
was  not  so  liable  to  get  organic  destruction  within  the 
joint  itself;  therefore  the  consequent  anchylosis  following 
upon  this  condition  was  considerably  reduced,  and  a  better 
future  usefulness  of  the  patient  assured.  This  was,  no 
doubt,  due  to  the  fact  that  the  synovia  was  laden  with 
active  bacteria  or  their  products,  which  in  themselves 
possess  an  irritating  effect  upon  the  synovial  membranes 
and  articular  surfaces,  causing  cell  proliferation,  cell  death, 
rarefying  ostitis,  anchylosis  of  the  joint,  and  consequent 
destruction  of  its  free-moving  capacity. 

In  those  cases  where  the  outset  of  the  disease  takes 
place  soon  after  parturition,  running  a  rapid  course  with 
great  prostration  of  the  patient,  giving  strong  suggestions 
of  a  septicaemic  condition,  little  by  way  of  treatment  can 
be  done.  Lignieres  inclines  to  the  belief  this  condition 
is  due  to  the  coccobacillus,  in  which  case  his  vaccine 
(polyvalent)  may  do  good;  but  of  this  Ave  have  no  ex- 
perience. 

Considering  the  ubiquity  of  the  coccobacillus,  it  is  just 
possible  that  this  bacterium  is  the  primary  factor  in  the 
causation  of  joint-evil,  and  that  other  bacteria  present  are 
more  or  less  accidental  and  secondary.  In  like  manner,  it 
appears,  human  tuberculosis  claims  its  many  victims,  not  so 
much  to  the  specific  bacilli  of  tubercle  itself,  but  to  the 
secondary  invasion    of   streptococci,  staphylococci,  Micro- 


VACCINE-THERAPY  109 

coccus  tetragenus,  etc.  The  same  also  may  be  said  of 
influenza,  distemper,  and  strangles,  as  we  shall  see  later. 

If  one  aspirates  an  infected  joint,  in  our  experience,  in 
the  early  stages,  centrifugalizes  the  fluid,  and  with  the 
sediment  inoculates  a  culture-tube,  no  growth  may  follow, 
and  this  rather  suggests  the  fact  that  the  bacteria  do  not 
enter  the  capsule  of  the  joint  at  the  outset,  but  rather 
remain  in  the  blood  or  lymph  stream,  toxins  only  filtering 
through,  causing  turbidity  of  the  otherwise  clear  synovia ; 
but  later  on,  owing  to  disintegration  of  the  vessel  walls, 
they  are  liberated,  and  find  their  way  into  the  joint  cavity. 
Be  this  as  it  may,  we  have  found  the  longer  standing  a 
pyaamic  arthritic  is,  the  greater  seems  to  be  the  degree 
of  bacterial  invasion,  and  vice  versa. 

A  careful  examination  of  the  navel  cord  should  be  made. 
The  purulent  matter  should  not  be  taken  from  the  point  of 
the  navel.  If  the  cord  is  fairly  long,  the  tip  must  be  cut 
off,  the  haemorrhage  stopped,  and  a  sterile  small  curette 
inserted  an  inch  or  two  into  the  canal,  and  a  scraping 
taken  from  the  side  of  its  wall.  This  should  be  examined 
and  incubated. 

As  already  pointed  out,  mixed  infection  is  rather  the 
rule,  and  it  therefore  follows,  each  case,  to  obtain  the 
maximum  of  success,  should  be  treated  upon  its  own 
merits. 

The  most  common  bacteria  the  writer  has  found  are — 
staphylococci  (albus  and  aureus) ;  streptococci ;  in  one 
case  the  B.  pyocyaneus,  in  another  a  microbe  resembling 
the  pneumococcus,  and  showing  its  characteristics;  and 
lastly  we  have  isolated  the  B.  coli  communis  from  the  navel. 
Considering  how  readily  the  navel  cord  comes  in  contact 
with  faBcal  matter,  it  is  really  a  wonder  one  does  not  see 
more  cases  of  B.  coli  infection,  although  such  may  be  so, 
and  have  been  overlooked  in  practice. 

It  is  surprising  to  find  how  tolerant  young  foals  are  to 
big  doses  of  vaccine.  The  writer  remembers  the  first  two 
or  three   cases  he  injected,  feeling   rather    anxious  as  to 


110  CLINICAL  BACTERIOLOGY  AND 

the  results,  but  in  course  of  time  did  not  hesitate  to  give 
a  dose  of  500,000,000  devitalized  staphylococci  to  a  week- 
old  foal.  Of  course,  it  should  be  pointed  out,  through 
some  idiosyncrasies  of  the  patient  itself,  vaccines  act  less 
seriously  in  some  than  in  others;  and  that  the  vaccines 
seem  to  sensitize  more  acutely  some,  while  others  are  more 
refractory  to  their  effects.  One  must  remember  also  some 
vaccines  possess  in  a  pronounced  degree  the  power  of 
disturbing  the  metabolism  more  than  others. 

As  soon  as  we  are  called  in  to  a  case  of  joint-evil,  we  give 
an  initial  dose  of  the  following  stock  vaccines, 

Streptococci      50,000,000 

Staphylococci 100,000,000 

B.coli 50,000,000 

B.  jyyocyan&us 50,000,000 

and  watch  the  clinical  symptoms. 

We  then  examine  a  scraping  from  the  navel,  aspirate 
the  joint  or  joints,  if  more  than  one  is  involved,  centri- 
fugalize  and  examine  the  deposit,  cultivate  the  bacteria 
present  on  suitable  media,  and  make  the  necessary  vaccine. 
Where  a  joint  is  distended  with  fluid,  we  always  insist  on 
removing  it;  this  procedure  gives  the  animal  a  better 
chance  to  recover,  and  the  after-sequelae,  as  has  already 
been  pointed  out,  in  the  joint  itself  are  not  likely  to  be  so 
serious. 

If  the  patient  is  not  making  satisfactory  progress,  and 
should  a  streptococcus  be  present,  a  combined  vaccine  and 
serum  gives  good  results,  and  should  most  certainly  be 
tried. 

Septicemic  Arthritis  in  Cattle. 

This  disease  is  only  seen  in  cows  after  parturition,  and, 
in  the  writer's  experience,  in  complicated  parturitions, 
either  where  the  genital  organs  have  been  injured  during 
delivery,  or  where  the  placenta  or  part  of  the  placenta 
has  been  retained  in  utero :  in  short,  the  condition  is  due 


VACCINE-THERAPY  1 1 1 

to  a  septic  metritis  developing  into  a  septicaemia,  and  lead- 
ing up  to  a  specific  arthritis.  One  or  more  joints  may  be 
involved.  The  stifle,  by  reason  of  its  large  capacity,  and 
probably  its  nearness  to  the  centre  of  infection,  is,  in  the 
writer's  experience,  particularly  singled  out  for  bacterial 
invasion. 

Of  course  one  sees  many  cases  in  practice  where  the 
placenta  is  retained  for  a  long  period,  and  very  septic, 
germ-laden  discharges  are  evacuated,  perhaps  for  weeks, 
and  yet  no  arthritic  lesions  develop,  nor  may  we  find  any 
indications  whatever  of  septic  infection  or  intoxication ; 
while,  on  the  contrary,  the  placenta  may  be  retained  only 
a  few  days,  or  there  may  only  be  slight  bruising  of  the 
genital  passage,  and  septicseniia  and  septic  arthritis  de- 
velop. We  can  only  explain  these  extreme  conditions  by 
stating  in  the  one  case  the  bacteriotropic  forces  were  in 
themselves  capable  of  acting  as  a  resisting  barrier  against 
a  systemic  bacterial  invasion,  and  in  the  other  they  were 
not.  So  far  as  our  present  knowledge  goes,  it  would 
appear  there  exists  in  the  blood-serum  a  specific  opsonin 
for  all  or  nearly  all  pathogenic  bacteria ;  thus,  we  may  have 
an  antistaphylococcal  opsonin,  an  antistreptococcal  op- 
sonin, an  anticolic  opsonin,  and  so  on.  The  quantity  of 
opsonin  differs  in  various  animals,  and  even  in  the  same 
animal  at  different  times ;  therefore  an  animal  may  be,  and 
is,  more  resistant  to  bacterial  invasion  at  one  time  than  at 
another. 

This  variation  is  dependent  upon  many  and  varied  causes, 
which  in  themselves  are  chemically  complex  and  difficult  to 
understand. 

But  if  we  do  not  know  why  these  opsonins  and  other 
antibodies  diminish  in  quantity  as  they  do,  we  do  know 
how  to  raise  them  to  a  standard  of  proficiency,  and  so 
prepare  the  antibodies  to  clear  the  system  of  offensive 
bacteria — i.e.,  by  using  a  suitable  vaccine  or  serum,  or  both 
combined. 

Where    an   animal    has    a   high    temperature,  is   off   its 


112  CLINICAL  BACTERIOLOGY  AND 

appetite,  with  more  than  one  joint  involved,  and  in  addi- 
tion, perhaps,  pneumonic  lesions  are  diagnosed,  an  acute 
septicemia  has  become  established,  and  little  hope  of 
recovery  is  likely  to  follow,  in  such  a  case  as  this  vaccine- 
therapy  will  avail  but  little;  but  should  the  disease  be 
running  a  slow  and  insidious  course,  and  the  circulation 
comparatively  free  from  bacteria  or  their  toxins,  much  can 
be  done  by  using  suitable  vaccines. 

The  writer  has  isolated  from  the  joints  of  cattle  a  strepto- 
coccus similar  to  those  obtained  from  cows  infected  with 
endometritis,  and  in  one  case  a  streptococcus  has  been 
obtained  from  the  blood  of  a  case  suffering  from  septicaemia 
and  metritis  with  arthritic  complications. 

We  do  not  get  the  same  bursal  distensions  in  cows  as  is 
seen  in  foals  with  pyemic  arthritis,  and  this  is  no  doubt 
due  to  the  fact  that  in  the  young  animal  the  bursae  are 
more  elastic  and  distend  more  freely  on  pressure  than  in 
older  animals.  We  therefore  have  not  the  same  necessity, 
nor  do  we  derive  the  same  benefit  from  aspirating  the  joint 
of  the  cow  that  we  obtain  by  so  doing  in  the  foal. 

In  treating  septicemic  arthritis  in  cattle,  we  must 
endeavour  to  destroy  the  bacteria  at  the  source  of  infec- 
tion. If  it  is  a  retained  piece  of  placenta  it  must  be 
removed ;  if  it  is  a  septic  uterine  wound  it  must  be  rendered 
aseptic.  Then  we  must  see  the  blood  is  kept  as  free  as 
possible  from  bacteria  or  their  products,  in  assisting  Nature 
to  get  rid  of  effete  material  by  keeping  the  eliminative 
organs  free.  And,  lastly,  we  must  help  her  to  elaborate 
her  protective  forces  by  using  appropriate  vaccines. 

When  one  has  a  case  showing  symptoms  of  infection  or 
intoxication,  with  premonitory  joint  lesions,  or  when  a 
placenta,  even,  has  been  retained,  and  it  appears  to  be  taking 
a  serious  turn,  one  is  fully  justified  in  giving  as  a  prophy- 
lactic a  polyvalent  staphylococcal  and  streptococcal  stock 
vaccine,  and,  if  preferred,  combined  with  an  antistrepto- 
coccal  serum.  But  to  be  of  any  real  service  the  bacterial 
emulsion    should  be  taken  from    a    culture    derived   from 


VACCINE-THERAPY  113 

the    genital  organ  or  joint   of    a  cow   suffering  from  the 
disease. 

Where  the  immunizer  has  not  a  stock  vaccine  at  hand, 
a  culture  should  be  made  from  the  uterine  discharge,  and 
also,  if  possible,  from  the  joint.  This  latter  may  be  difficult 
and  sometimes  impossible  to  do,  by  reason  of  the  fact  the 
bacteria  may  not  be  present  in  the  synovia  at  all,  as  already 
explained  in  the  case  of  young  foals. 

If,  however,  the  animal  dies,  one  can  usually  obtain 
active  bacteria  from  the  neighbourhood  of  the  joint,  and 
invariably  the  infection  is  a  mixed  one. 

Cattle  are  very  tolerant  to  big  doses  of  vaccine;  the 
writer  does  not  hesitate  to  give  1,000,000,000  staphylococci 
and  500,000,000  streptococci  as  an  initial  dose,  and  doubling 
those  doses  in  five  days  if  necessary. 

We  have  found  cases  of  septicemic  arthritis  improve 
more  rapidly  under  vaccine-therapy,  lay  on  flesh  more 
quickly,  and  not  so  liable  to  suffer  from  subsequent  and 
permanent  lameness,  thus  giving  better  results  than  can  be 
obtained  by  any  other  mode  of  treatment. 

Moreover,  by  adopting  prophylactic  and  curative  vaccines 
lives  have  been  saved  in  our  hands  which  in  all  probability 
would  have  died  from  auto-infection  or  intoxication,  or 
both. 

It  is  quite  feasible  to  suspect  the  B.  coli  communis 
may  attack  joints,  in  a  manner  similar  to  streptococci,  from 
the  womb,  and  the  immunizer  should  be  on  the  lookout  for 
such  an  invasion. 


CHAPTER  XIV 

BACTERIAL  DISEASES  AFFECTING  THE 
ABDOMINAL  ORGANS 

Disease.  Cause. 

Enteric  :  Diarrhoea  and  dysentery      B.  coli  group,  B.  pijocyaneus,  B. 

jiroteus,  streptococci,  B.  tuber- 
culosis.    Johne's  bacillus. 

Peritoneal:  Peritonitis  B.  tuberculosis,  B.  coli,  staphy- 
lococci, streptococci. 

Nephritic:  Pyelonephritis,  cystitis      Pneumococcus,     B.    coli     group, 

streptococci,  staphylococci. 

Uterine :  Endometritis     ...         ...      Pneumococcus,     B.    coli     group, 

staphylococci,  streptococci. 

Diarrhoea  and  Dysentery. 

As  it  is  only  natural  to  expect,  the  intestines,  by  reason 
of  their  functions,  especially  the  lower  bowels,  are  infected 
by  a  long  list  of  bacteria,  some  pathogenic,  many  non- 
pathogenic. Some  of  the  non-pathogenic  bacteria,  however, 
adopt  a  disease-producing  role  under  certain  circumstances 
only,  of  which  several  of  the  B.  coli  group  are  examples. 

Young  animals  are  particularly  prone  to  bacterial  intes- 
tinal invasion,  and  the  reason  probably  is  that  milk  is  up 
to  a  certain  period  their  staple  diet,  and  milk  is  an  excellent 
medium  for  bacterial  growth  and  transmission.  Moreover, 
the  intestinal  mucosa  is  less  resistant  in  the  young  animal 
than  in  the  old.  It  is  just  possible,  also,  as  an  animal  gets 
older,  repeated  mild  auto-infections  from  the  intestinal 
mucosae  are  continually  taking  place,  conferring  upon  the 
subject  a  degree  of  resistance,  which  with  age  becomes 
merged  into  an  immunity. 


VACCINE-THERAPY  115 

Where  one  gets  isolated  mild  attacks  of  diarrhoea  in 
young  animals,  the  ordinary  medicinal  treatment  answers 
all  practical  purposes ;  but  when  an  attack  affects  a  large 
number  of  animals,  with  a  high  death-rate,  prophylactic 
and  curative  measures  of  the  most  up-to-date  kind  must  be 
adopted. 

It  is  in  cases  such  as  these  that  vaccine-therapy  should 
be  used.  In  young  animals  the  alimentary  canal  becomes 
infected  from  two  distinct  sources — i.e.,  oral  and  umbilical. 
To  what  source  one  can  attribute  the  greater  number  of 
cases  it  is  difficult  to  hazard  an  opinion,  but  the  fact  that 
both  are  capable  channels  of  infection  the  practitioner 
should,  in  investigating  any  outbreak,  always  bear  in 
mind. 

As  already  stated,  a  large  variety  of  bacteria  are  to  be 
found  in  the  intestinal  canal,  and  it  is  therefore  of  the 
first  importance  that  one  should  ascertain  what  bacteria 
are  taking  on  a  pathogenic  role. 

But  this  is  not  all.  Having  isolated  from  scrapings  or 
excreta,  or  both,  a  variety  of  bacteria,  one  has  to  decide 
which  is  the  most  capable  of  producing  the  disease  in 
question.  Of  course,  this  delicate  point  confronts  the 
immunizer  in  all  cases  of  mixed  infections,  more  or  less, 
and  it  must  largely  be  left  to  his  own  individual  knowledge 
upon  which  data  he  bases  his  conclusions,  provided  they 
are  always  in  keeping  with  the  general  laws  of  bacteri- 
ology. The  B.  coli  group  appear  under  certain  circum- 
stances to  develop  very  potent  pathogenic  qualities,  and 
it  is  probable  they  are  the  forerunners  of  many  other 
grave  bacterial  intestinal  invasions.  Some  authorities 
believe  the  pasteurella  group  play  a  most  important 
part. 

In  addition  to  these  we  have  the  B.  pyocyaneus,  B.  pro- 
teus,  and  streptococcus ;  while  we  find  such  specific  in- 
vasions from  B.  tuberculosis,  B.  Johne,  and  Streptothrix 
actinomyces. 

In  the    condition    known   as  "  white  scour "   in    calves, 


116  CLINICAL  BACTERIOLOGY  AND 

we  have  obtained  good  results  from  the  early  injection  of 
a  stock  B.  coli  vaccine,  usually  beginning  with  a  dose  of 
25,000,000  in  young  calves.  It  is  advisable,  where  an  autog- 
enous vaccine  is  not  used,  a  polyvalent  one  should  take  its 
place. 

A  second  injection  of  50,000,000  should  be  given  three 
or  four  days  later,  and  the  conditions  noted. 

In  addition  to  vaccines,  in  many  bacterial  intestinal  in- 
vasions we  have  had  excellent  results  from  the  early  use 
and  repeated  administration  of  boric  acid. 

Johne's  Disease  in  Cattle. 

It  may  not  be  out  of  place  here  to  mention  a  rather 
common  bacterial  invasion  of  the  intestines  of  (usually) 
adult  cattle  in  certain  districts  by  an  acid-fast  bacterium 
resembling  in  many  ways  the  bacillus  of  tubercle,  and 
named,  after  the  discoverer,  Johne's  bacillus. 

Practitioners  in  days  gone  by  considered  Johne's  disease, 
as  seen  in  cattle,  to  be  a  form  of  enteric  tuberculosis. 

The  specific  bacillus  can  be  easily  isolated,  stained,  and 
demonstrated  under  the  microscope,  TV  oil  immersion,  from 
scrapings  made  from  the  intestines.  It  is,  however,  a  very 
shy  microbe  to  cultivate  artificially  in  the  laboratory  upon 
the  usual  media ;  but,  thanks  to  the  well-known  research 
work  of  Twort  and  Ingram,  a  medium  has  been  prepared 
to  suit  the  growing  tastes  of  this  bacterium.  These  two 
workers  found  by  cultivating  and  killing  the  Timothy- 
grass  bacillus  (B.  phlei),  and  mixing  the  result  with  the 
nutrient  media,  the  bacillus  of  Johne  grew  most  success- 
fully, the  reason  they  give  being  that  probably  the 
Timothy -grass  bacillus  is  the  "  wild  ancestor"  of  the 
Johne  bacillus. 

They  have  made  a  vaccine,  as  above  indicated,  which 
they  assert  has  a  diagnostic  value  towards  Johne's  disease 
equal  to  that  which  tuberculin  has  to  tuberculosis,  and 
mallein  has  to  glanders ;  and  they  are  careful  to  point 
out  that  the  maximum   rise    of   temperature    is    expected 


VACCINE-THERAPY  1 1 7 

about  the  tenth  hour,  which,  of  course,  is  earlier  than  with 
mallein  or  tuberculin. 

They  are  hopeful  also  the  vaccine  may  possess  curative 
virtues  as  well,  but  on  this  point  further  investigation 
appears  to  be  necessary. 

Peritonitis. 

The  peritoneum  forms  in  itself  an  admirable  breeding- 
ground  for  many  bacteria,  and  probably  none  grow  more 
luxuriantly  than  the  bacillus  of  tubercle,  especially  on  the 
bovine  peritoneum. 

Bacteria  gain  entrance  to  the  peritoneum  in  a  variety  of 
ways. 

The  B.  coli,  existing  as  it  does  in  the  intestines,  often 
finds  it  way  into  the  peritoneal  cavity  through  abrasions 
in  the  intestinal  wall,  commonly  due  to  the  bursting  of  an 
abscess,  the  penetrating  effects  of  intestinal  parasites, 
migration  from  the  womb  in  endometritis,  injuries  from 
without,  etc.  When  they  escape  from  the  intestines  they 
multiply  with  great  rapidity,  forming  toxins  which,  by 
reason  of  the  physiological  functions  of  the  peritoneal 
membrane,  become  readily  absorbed  into  the  circulation, 
producing  intoxication,  infection,  and  death.  It  is  obvious 
in  these  very  acute  cases  vaccine-therapy  will  avail  little, 
and,  for  that  matter,  any  other  system  of  treatment. 

When,  however,  the  condition  becomes  circumscribed 
and  subacute  or  chronic  in  contra-distinction  to  diffused 
and  acute,  if  the  case  can  be  so  diagnosed,  much  good 
will  follow  vaccine  treatment,  and  the  peritoneum  par- 
ticularly lends  itself  to  such  therapy  by  reason  of  the  fact 
that  it  is  continually  bathed  by  a  process  of  osmosis  with 
the  blood-serum,  which  contains  the  protective  elements  of 
the  blood.  Of  course,  here  we  find  limitations  compared 
with  human  medicine  and  surgery,  but  it  sometimes 
happens,  particularly  in  the  smaller  animals,  abdominal 
surgery  is  performed,  when  we  have  developed  a  plastic 
entero-peritonitis  due  to  the  B.  coli  perhaps.     The  system 


118  CLINICAL  BACTERIOLOGY  AND 

can  be  considerably  strengthened  and  fortified  if  a  suitable 
vaccine  is  administered  in  addition  to  local  treatment  and 
proper  drainage. 

The  B.  coli  may  gain  a  footing  in  the  peritoneum,  espe- 
cially in  cows  after  parturition,  through  the  womb,  seen  in 
such  cases  as  metroperitonitis.  Here,  again,  where  we 
suspect  such  a  condition,  much  can  be  done  by  vaccines. 

Streptococci  may  infect  the  peritoneum  through  strangles 
attack  running  an  irregular  course,  and  also  from  the  womb 
in  mares  and  cows. 

The  cocci  may  locate  themselves  in  a  mesenteric  gland, 
which  in  time  bursts,  the  bacteria  are  liberated,  and  peri- 
tonitis sets  in.  Here,  again,  if  the  condition  is  diffused 
and  acute,  little  good  will  follow  any  line  of  treatment, 
but  should  it  be  limited,  and  not  too  severe,  much  benefit 
will  follow  the  injection  of  20  c.c.  antistreptococcic  serum 
and  a  vaccine  of  500,000,000  devitalized  streptococci  to  a 
horse,  repeated  in  three  to  five  days  in  a  double  quantity. 

It  is  more  than  probable  that  the  majority  of  the  peri- 
toneal inflammations  we  see  in  practice  are  of  bacterial 
origin,  except  those  cases  due  to  traumatism;  and,  of  course, 
they  also  soon  become  organismally  contaminated,  if  they 
are  not  already  so. 

Moreover,  those  acute  cases  of  muco-enteritis  which  we 
so  often  see  in  equine  practice  affecting  high-conditioned, 
grossly-fed  animals  are  all  probably  due,  either  primarily 
or  secondarily,  to  bacterial  invasion. 

Pyelonephritis. 

Diseases  in  general  of  the  urinary  system  have,  on  the 
whole,  been  seriously  neglected  by  practitioners  of  com- 
parative medicine.  This  is  partly  accounted  for  by  the 
fact  that  even  grave  nephritic  conditions  may  exist,  and 
give  no  positive  indications  of  their  existence  to  the  clinical 
observer.  More  important,  perhaps,  is  the  undeniable  fact 
that  clinical  investigations  are  not  as  thorough  as  they 
might  be,  nor  are  they  as  often  applied  as  they  should  be. 


VACCINE-THERAPY  119 

We  are  all  aware  of  cases  which  from  time  to  time  come 
under  observation  where  the  kidneys  are  reduced  to  a 
mass  of  almost  useless  pulp,  and  yet  during  life  we  never 
suspected  or  diagnosed  nephritic  trouble.  Had  a  careful 
examination  of  the  urine  in  such  a  case  been  carried  out, 
our  diagnosis  and  treatment  might  have  been  different  and 
the  results  perhaps  more  satisfactory. 

From  a  fairly  extensive  bacteriological  examination  of 
the  urine,  especially  in  equines,  we  have  been  struck  with 
the  great  susceptibility  of  the  urinary  system  to  bacterial 
invasions.  And  this  is  not  surprising,  considering  the 
anatomical  arrangements  and  excretory  functions  of  the 
urinary  system. 

Bacterial  infection  can  take  place  from  two  sources : 
(1)  Infection  internally  from  and  through  the  blood- 
stream j  (2  infection  externally  from  the  vulva  in  the 
female,  which  is  rather  common,  or  in  the  male  through 
the  urethra,  which  is  very  uncommon.  Considering, 
therefore,  the  eliminative  functions  of  the  urinary  system, 
and  its  anatomical  arrangements  and  position,  bacterial 
invasions  must  needs  be  nearly  always  mixed.  The 
most  common  bacteria  found  in  practice  are  streptococci, 
staphylococci,  B.  pyocyaneus,  B.  coli  communis,  B.  tuber- 
culosis. As  an  illustration  of  bacterial  infection  through 
the  blood-stream,  one  may  cite  metastatic  strangles,  where 
the  streptococci  become  entangled  in  the  parenchyma  of 
the  kidneys  following  upon  metastasis. 

External  infection  is  a  condition  which  is  not  to  be 
wondered  at,  considering  the  unhygienic  state  of  the  animal 
rectum,  vulva,  buttocks,  tail,  etc.,  the  close  relationship  the 
vulva  has  with  the  rectum,  and,  lastly,  the  nearness  of  the 
meatus  urinarius  to  the  vulval  opening. 

In  pregnant  animals,  following  upon  delivery  or  abortion, 
one  sometimes  finds  cystic  and  nephritic  bacterial  invasions, 
as  a  result  of  extension  from  the  uterus,  vagina,  or  vulva. 

When  one  suspects  disease  or  derangement  of  the  urinary 
system,  a  careful  examination  of  the  urine  should  be  made. 


120  CLINICAL  BACTERIOLOGY  AND 

A  sterile  catheter  is  passed  into  the  bladder,  and,  after 
allowing  a  small  proportion  of  the  urine  to  flow  away,  the 
remainder  should  be  collected  in  a  sterile  jar  and  sealed 
for  examination. 

The  urine  thus  collected  is  centrifugalized,  and  the 
resultant  deposit  examined  after  removal  of  the  supernatant 
fluid  by  pipette  or  pouring. 

A  hanging-drop  preparation  should  be  made  of  the 
sediment  and  carefully  examined  under  a  \  and  -^  oil 
immersion.  One  will  be  able  thus  to  detect  bacteria  if 
present,  and  note  if  they  are  motile,  if  single  or  in  pairs, 
or  in  chains  or  bundles,  etc.  Films  should  now  be  made 
in  the  ordinary  manner  and  stained.  If  any  difficulty 
is  encountered  in  fixing  the  film  upon  the  slide,  a  drop 
of  one's  own  blood  from  a  finger-prick  and  well  mixed 
with  the  urine  will  supply  sufficient  albumin  to  the  field 
to  fix  the  specimen  when  heat  or  other  fixing  agents  are 
applied. 

When  a  culture  is  desired,  the  collection  of  the  urine 
and  the  inoculation  of  culture  media  requires  the  most 
careful  application  to  prevent  risk  of  outside  contamination. 

Where  the  kidneys  are  extensively  invaded  by  bacteria, 
the  process  of  functional  elimination  in  these  organs 
decreases,  and  with  destructive  changes  going  on  in  the 
organs  themselves,  bacteria  and  tissue  cells  are  continually 
passing  along  the  urinary  tract.  Some  of  these  pathogenic 
organisms  settle  in  the  mucosa,  causing  cystitis  and 
urethritis;  others  are  expelled  with  the  urine;  and  super- 
added in  advanced  cases  one  sees  typical  uremic  symptoms. 
Drugs  in  such  cases  give  little  relief,  although  alkalies, 
urotropin,  and  the  volatile  oils,  may  give  temporary 
benefits. 

Having  ascertained  the  causative  bacteria,  a  suitable 
vaccine  should  be  made  in  the  usual  manner,  and  the 
phenomena  carefully  watched. 

If  uraemia  has  already  set  in,  much  assistance  can  be 
given  to  Nature  by  lowering  the  blood-pressure  and  stimu- 


VACCINE-THERAPY  121 

lating  the  other  excretory  organs — i.e.,  the  bowels  and  skin. 
The  subcutaneous  or  intravenous  injection  of  normal  saline 
solution  assists  the  system  also  in  the  removal  of  an  ever- 
accumulating  effete  deposit. 

Endometritis. 

Bacterial  invasions  of  the  womb  in  the  lower  animals  are 
of  rather  common  occurrence,  due  largely  to  contamination 
at  the  time  of  delivery  on  the  part  of  unskilled  attendants, 
and  also  by  the  retention  of  the  placenta. 

One  also  sees  many  cases  following  upon  difficult  par- 
turition, through  injuries  caused  to  the  mucosa  either  by 
the  foetus  itself  during  the  process  of  delivery,  the  acci- 
dental slipping  of  instruments,  and,  above  all,  the  want  of 
complete  asepsis  on  the  part  of  the  operator. 

Some  species  of  animals  are  more  tolerant  to  the  patho- 
genic influences  of  bacteria  growing  in  the  generative 
female  organ  than  others.  The  cow,  for  example,  is  prob- 
ably the  most  tolerant  of  all  the  domestic  animals,  while 
the  mare  is  the  least. 

Why  this  is  so  is  somewhat  difficult  to  explain,  but  it  is 
more  than  probable,  in  the  light  of  our  present  know- 
ledge, that  those  animals  which  are  most  resistant  are 
endowed  by  Nature  with  the  greatest  supply  of  bacterio- 
tropic  elements.  It  is  a  well-known  fact  that  the  uterine 
fluids  at  the  time  of  and  after  parturition  possess  powerful 
bacteria-destroying  properties,  and  it  may  be  due  to  those 
strong  bactericidal  qualities  also,  the  possession  of  which 
is  greater  in  some  species  of  animals  than  in  others,  that 
we  see  in  practice  such  variable  degrees  of  resistance 
versus  susceptibility. 

As  already  pointed  out,  uterine  bacterial  invasion  in 
practice  takes  place  incidentally  to  parturition  in  some 
form  or  another,  and,  indeed,  it  would  seem  almost  a 
physical  impossibility  for  the  womb  to  become  invaded  in 
animals  at  any  other  period,  for  during  a  process  of 
quiescence  of  that  organ  it  is  hermetically  sealed,  and  it 


122  CLINICAL  BACTERIOLOGY  AND 

would  only  be  the  result  of  some  mechanical  injury 
from  without  that  bacteria  could  reach  the  organ. 
Abortion  is  another  prolific  source  of  endometritis  and 
bacterial  invasions,  owing  to  the  fact  of  the  abortion 
bacillus  appearing  to  exert  a  lowering  effect  upon  the 
protective  elements  of  the  womb,  and  so  predisposing  it  in 
such  a  manner  to  the  easy  development  of  secondary 
infections.  The  womb  also  is  prevented  from  closing 
naturally  in  many  cases  owing  to  a  retained  placenta,  which 
putrefies,  giving  an  increased  impetus  to  further  bacterial 
invasions.  And  so  we  find  after  abortions,  that  bacterial 
infections  are  nearly  always  mixed. 

Some  of  these  infections,  either  due  to  the  great  activity 
of  the  bacteria  themselves  and  the  potency  of  their  toxins, 
or  the  weakened  bactericidal  power  of  the  animal  itself, 
run  a  very  rapid  and  highly  fatal  course,  death  taking  place 
in  a  few  hours  from  septic  infection,  or  even  intoxication. 
In  cases  such  as  these  the  vaccine-therapist  can  do  but 
little. 

When  the  condition  runs  a  more  subacute  course,  much 
can  be  done,  however,  by  using  suitable  vaccines. 

Where  the  infection  is  caused  by  septic  hands,  slips, 
instruments,  etc.,  during  the  course  of  a  difficult  parturition, 
it  is  invariably  a  mixed  one,  the  streptococci,  staphylococci, 
B.  coli  communis,  etc.,  being  the  more  common  offenders. 

To  ascertain  the  causative  bacteria,  we  make  it  a  practice 
to  thoroughly  wash  the  vulva  and  buttocks  with  soap 
and  an  antiseptic  ;  we  then  insert  the  hand  and  arm, 
carrying  into  the  womb  a  sterile  test-tube  which  has  been 
cut  down  to  2  inches  in  length.  By  drawing  the  mouth 
along  the  mucosa  close  to  the  neck  of  the  uterus,  we  obtain 
sufficient  materia  morbis  for  examination.  This  is  plugged 
with  a  sterile  rubber  cork,  and  marked  "No.  1."  In  like 
manner  another  tube  is  carried  in,  to  abstract  material  in 
the  body  of  the  womb,  and  marked  "  No.  2 ";  while  with 
a  third  tube  material  is  taken  as  far  forward  as  one's  hand 
will  reach,  and  marked  "  No.  3." 


VACCINE-THERAPY  123 

By  this  method  we  can  ascertain — 

1.  What  bacteria  we  are  fighting  against. 

2.  We  know  nearly  what  stage  of  infection  the  patient 
is  in  at  the  time  of  examination ;  that  is  to  say,  if  the  in- 
fection is  very  recent,  and  the  bacteria  have  not  invaded  the 
whole  of  the  womb,  material  from  No.  1  tube  will  contain 
more  bacteria  than  are  to  be  found  in  tubes  Nos.  2  and  3. 
On  the  contrary,  if  the  invasion  is  equally  diffused  over 
the  whole  organ,  the  three  tubes  should  be  practically 
alike. 

Of  course,  if  the  case  is  an  advanced  one  and  the  prac- 
titioner is  called  in  late,  the  womb  may  be  sufficiently 
closed  to  prevent  the  entrance  of  one's  hand  unless  ill- 
advised  force  is  used ;  but,  on  the  contrary,  the  fact  that 
the  womb  is  in  such  a  diseased  condition  militates  against 
its  natural  processes  of  contraction.  In  fact,  it  is  this 
contractile  power  the  practitioner  so  much  desires  to  be 
brought  about,  and  to  accelerate  the  condition  in  practice 
we  make  a  point  of  giving  full  doses  of  ergot,  thereby 
limiting  the  risk  of  extension  and  the  possible  development 
of  septic  infection  or  intoxication. 

To  ascertain  the  degree  of  infection  in  each  tube,  we 
place  an  equal  quantity  of  the  material  in  a  sterile  tube, 
and  add  a  given  quantity  of  sterile  salt  solution,  draw  off 
the  end  of  the  tube  in  the  blow-flame,  seal  it  up  as  already 
described,  and  mark  it  with  grease  pencil  "No.  1."  The 
same  process  is  gone  through  with  Nos.  2  and  3. 

These  are  well  shaken — each  for  a  given  time — to  break 
down  the  bacterial  bundles,  chains,  etc.  A  drop  of  the  fluid 
is  now  placed  on  the  end  of  a  slide,  spread  carefully  with 
a  Wright  spreader,  and  allowed  to  air  dry  and  marked 
"  No.  1."  The  same  method  is  adopted  with  Nos.  2  and  3. 
These  are  respectively  examined,  and  the  bacteria,  say  in 
half  a  dozen  squares  of  the  microscopic  field,  are  counted 
and  totalled,  and  each  slide  total  is  then  compared.  If 
the  films  are  equally  spread  in  each  case,  the  results  should 
be  pretty  accurate. 


124  CLINICAL  BACTERIOLOGY 

Having  ascertained  what  bacteria  are  present,  the  im- 
munizer  should  forthwith  make  an  autogenous  vaccine. 

In  our  experience  the  B.  coli  groups  are  the  most  common 
offenders,  and  they  sometimes  take  on  a  very  virulent  form. 
After  injecting  the  vaccine,  a  careful  note  of  the  clinical 
symptoms  should  be  made.  A  slight  rise  of  temperature 
may  take  place,  and  an  increase  of  the  uterine  discharge 
is  to  be  expected.  After  forty- eight  hours  this  should 
become  gradually  less,  the  patient  will  look  brighter,  the 
temperature  falls;  and  this  will  go  on  for  a  few  days, 
after  which  another  injection  should  be  given. 

Locally,  of  course,  much  can  be  done  to  assist  the 
vaccines. 

The  womb  should  be  irrigated  with  a  very  dilute  boric 
acid  solution,  to  which  is  added  citrate  of  soda  in  weak 
solution. 

The  temperature  of  the  douche  should  be  about  40°  C, 
and  gradually  increased  to  45°  C.  The  therapeutic  value 
of  this  is  apparent.  The  womb  is  a  highly  vascular  organ. 
By  applying  heat,  and  gradually  increasing  that  heat, 
dilatation  of  the  bloodvessels  takes  place,  more  blood 
reaches  the  organ,  increased  osmosis  follows,  and  so  long 
as  the  blood  in  such  a  state  can  be  kept  fluid,  as  it  can  be 
with  the  soda  citrate  in  solution,  so  long  will  an  increased 
flow  of  bacteriolytic  and  bacteriotropic  essentials  reach  the 
death-producing  bacteria,  to  the  benefit  of  the  patient. 

Needless  to  say,  these  uterine  douches  should  be  repeated 
at  intervals,  and  the  fluid  should  not  remain  in  the  womb 
longer  than  a  few  minutes,  it  being  either  pumped  or  siphoned 
off  if  the  contractile  powers  of  the  womb  itself  are  in 
abeyance,  or  the  patient  is  too  weak  to  make  an  effort. 

When  streptococci  are  present,  a  streptococcal  serum 
should  also  be  used,  20  c.c.  being  injected  daily  for  several 
days  with  increasing  doses;  but  note  should  be  made 
of  the  fact  that  sera  only  obtained  from  a  bovine  animal 
should  be  used,  such  animal  having  been  immunized  by  the 
uterine  streptococcus  for  preference. 


CHAPTER  XV 

BACTERIAL  DISEASES  AFFECTING  THE 
CIRCULATORY  SYSTEM 

Disease.  Cause. 

Septicaemia     Streptococcus  septus,  staphylococci,  pneumo- 

cocci,  B.  coli. 
Pyeeruia  Streptococcus  septus,  staphylococci,  pneumo- 

cocci,  B.  coli. 
Malignant  oedema     ...      Bacillus  of  malignant  cedema. 

Quarter-evil Bacillus  of  quarter-evil. 

Anthrax  Bacillus  of  anthrax. 

Septicaemia. 

The  diseases  under  this  heading  are  more  commonly  seen 
in  veterinary  practice  than  in  human  medicine,  and  the 
reason  is  not  far  to  seek.  The  predisposing  causes  to  all 
septicemias,  speaking  generally,  are  neglect  and  unhealthy 
surroundings. 

Septicemia  appears  as  two  distinct  conditions — i.e.,  (1) 
septic  infection,  when  the  blood  itself  is  charged  with 
living  bacteria.  If  such  blood  is  injected  into  the  circu- 
lation of  a  healthy  animal  a  similar  disease  will  be  produced 
in  that  animal ;  and  (2)  septic  intoxication,  when  the  blood 
is  charged  with  poisons  emanating  from  bacteria  located  in 
some  centre  in  the  body.  If  such  blood  is  injected  into  a 
healthy  animal,  no  reproduction  of  that  disease  takes  place, 
thereby  distinguishing  septic  infection  from  septic  intoxica- 
tion. Although  we,  therefore,  recognize  in  practice  these 
two  conditions  as  separate  ones,  the  clinician  should  always 
remember  that  an  intoxication  may  become  an  infection 


126  CLINICAL  BACTERIOLOGY  AND 

at  any  moment,  and  as  practitioners  we,  therefore,  should 
be  on  the  lookout  for  such  developments. 

In  septic  infection  the  bacteria  gain  entrance  to  the 
blood-stream  or  lymphatics  usually  through  a  solution  of 
continuity  of  the  skin  or  mucous  membrane,  and  there 
must  be  a  tendency  towards  some  defective  bactericidal 
power  on  the  part  of  the  patient,  and  probably  phago- 
cytosic  inertia ;  for,  as  one  knows,  so  soon  as  a  local  centre 
becomes  infected,  a  ring  of  active  cells  is  formed  round 
that  area,  thereby  imprisoning  the  pathogenic  elements. 

Should  this  ring  remain  intact,  the  most  that  will  follow 
will  be  osmosis  of  toxins  through  the  barrier,  which  may  get 
into  the  circulation  and  set  up  septic  intoxication,  with 
constitutional  disturbance,  if  the  toxin  is  virulent  enough 
and  sufficient  has  been  absorbed.  If,  however,  the  wall 
gives  way  through  any  defective  effort  of  Nature  on 
the  one  hand,  and  extreme  activity  and  virulence  of  the 
bacteria  on  the  other,  septic  infection  follows. 

It  is  notorious  the  number  of  small  punctured  but  deep- 
seated  wounds  which  act  as  a  starting-point  in  the  develop- 
ment of  a  generalized  septicemia  compared  with  superficial 
and,  maybe,  more  extensive  lacerated  wounds.  This  is 
no  doubt  due  to  the  fact  that  small  punctured  wounds 
do  not  look  serious  in  the  mind  of  the  layman,  and 
the  owner  or  attendant  may  even  ignore  them  altogether ; 
whereas  a  superficial  and  an  extensive  wound  looks  ap- 
palling to  the  lay  observer,  and  he  sets  about  to  obtain 
advice  at  once.  And,  again,  a  punctured  wound  is  infected 
at  its  blind  end,  where  sunlight  and  ordinary  dressings  can 
scarcely  reach,  while  a  proper  temperature  for  bacterial 
growth  is  always  maintained,  and  the  result  is  rapid  bac- 
terial development,  with  grave  constitutional  phenomena. 

The  causative  bacteria  usually  found  in  wound  infection 
are  the  streptococci  and  staphylococci ;  and  after  castrations, 
particularly  in  lambs,  we  have  isolated  a  bacillus  answering 
the  description  of  the  B.  colt  communis — motile  and  Gram- 
negative,  etc. 


VACCINE-THERAPY  127 

Another  common  illustration  of  septic  infection  and 
intoxication  is  to  be  found  in  cows  after  parturition  in  that 
condition  known  as  "  septic  metritis/'  As  already  ex- 
plained under  the  heading  of  endometritis,  bacterial 
infection  takes  place  through  inoculation  caused  by  septic 
hands  on  the  part  of  the  operator,  slips,  instruments, 
injuries  after  parturition,  and  retained  placenta,  par- 
ticularly after  abortion. 

Here  the  B.  coli,  streptococci,  and  staphylococci  play  an 
important  role.  In  advanced  cases  of  this  kind  we  have 
invariably  noticed,  especially  in  heifers,  in  addition  to  the 
characteristic  septic  chocolate-coloured  discharge  from  the 
womb,  a  slimy  mucoid  discharge  from  the  nostrils,  accom- 
panied by  a  persistent  bronchial  cough.  From  this  dis- 
charge the  writer  has  isolated  streptococci  and  diplococci, 
and  has  been  struck  by  the  smallness  of  the  chains,  both 
from  the  bronchial  and  uterine  discharges,  compared  with, 
for  example,  the  streptococcal  chains  one  sees  in  strangles 
or  in  ordinary  wound  infections. 

We  cannot  think  the  pulmonary  streptococci  had  their 
primary  origin  in  the  uterus;  much  more  probable  is  it 
they  are  normal  inhabitants  of  the  pulmonary  mucosas,  and 
it  is  only  as  the  result  of  debility,  caused  by  the  uterine 
infection,  that  they  become  pathogenic  and  active.  Their 
presence  in  any  case  increases  the  liability  of  a  fatal  issue, 
and  the  practitioner  has  to  be  called  in  very  early  if  he 
hopes  to  bring  about  a  recovery. 

Such  a  case  requires  most  careful  attention.  A  stock 
vaccine  must  be  given  at  once,  preparatory  to  the  making 
of  a  suitable  autogenous  one ;  and  in  a  case  similar  to  the 
one  cited  above,  a  mixed  vaccine  composed  of  streptococci, 
B.  coli,  staphylococci,  and  diplococci  should  be  given.  If 
the  case  is  a  bad  one,  we  invariably  give  a  streptococcal 
serum  in  addition. 

Local  treatment,  of  course,  must  be  attended  to,  and 
here,  again,  we  must  most  strongly  deprecate  the  use  of 
strong  antiseptics.     In  the  case  of  a  deep  punctured  wound, 


128  CLINICAL  BACTERIOLOGY  AND 

it  must  be  laid  open  to  permit  of  free  drainage  and  the 
admission  of  dressings.  The  curette  is  a  most  useful  instru- 
ment for  the  removal  of  lodged  bacteria  on  the  edges  or 
at  the  blind  end  of  the  wound.  If  the  flow  of  lymph 
appears  deficient,  the  wound  should  be  irrigated  with 
citrate  of  soda  solution,  or  even  salt  and  water ;  and  when 
the  wound  appears  to  be  tardy  in  its  healing  process,  if 
located  in  the  limbs,  we  have  found  considerable  benefit 
from  the  application  of  Bier's  treatment. 

Where  one  is  dealing  with  septic  uterine  disease,  the 
womb  must  be  irrigated  and  a  similar  treatment  carried 
out  as  in  endometritis. 

Medicines  in  these  cases  can  be  of  little  avail.  Should, 
however,  the  temperature  be  high,  salicylic  acid  is  indi- 
cated, and  it  is  well  to  keep  the  eliminative  organs  active. 
Thus  the  bowels  should  be  kept  open  with  salines,  the 
kidneys  stimulated,  and  for  this  purpose  there  is  no  better 
drug  than  turpentine  ;  while  a  useful  diaphoretic  is  liq. 
amnion,  acet.  combined  with  salicylic  acid. 

Pyaemia. 

The  bacteria  involved  in  the  production  of  this  disease 
are  practically  similar  to  those  which  cause  septicaemia. 
The  characteristics,  however,  of  the  two  diseases  differ  in 
a  marked  degree.  In  pyaemia  multiple  abscesses  occurring 
in  various  centres  of  the  body,  as  the  result  of  metastasis, 
distinguish  this  disease  from  septicaemia. 

A  purulent  centre  may  be  the  means  of  producing 
pyaemia  through  the  protective  wall  giving  way,  the 
specific  bacteria  entering  the  blood  or  lymph  stream. 
Examples  of  this  are  to  be  found  in  strangles  in  the  horse, 
purulent  thrombophlebitis  in  foals  and  calves,  and  puer- 
peral pyaemia  in  the  mare  and  the  cow.  Where  abscess 
formations  are  found  in  the  internal  organs,  diagnosis  is 
not  always  easy,  and  the  disease  may  have  advanced  too 
far  for  treatment  to  be  of  any  avail.  Moreover,  these 
abscesses  are  generally  tensely  filled  with  pus,  which  pre- 


VACCINE-THERAPY  129 

vents  the  bactericidal  elements  from  reaching  the  septic 
foci.  If,  however,  the  abscesses  are  subcutaneous,  evacua- 
tion of  the  pus  is  easy  and  essential,  and  vaccine-therapy 
can  do  much  to  complete  a  cure. 

Where  the  pyaemia  is  a  sequel  to  strangles,  we  always  com- 
bine the  vaccine  with  an  antiserum ;  and  if  the  temperature 
is  very  high,  it  is  advisable  to  begin  with  a  minimum  dose 
at  the  outset.  If  the  temperature  does  not  rise  too  high 
during  the  negative  phase — in  three  or  five  days — we 
repeat  the  dose,  giving  a  double  quantity. 

Malignant  (Edema. 

This  condition  is  seen  in  several  species  of  the  domesticated 
animals,  and  is  due  to  a  wound  infection  by  the  bacillus  of 
malignant  oedema.  It  has  a  very  wide  distribution  in 
Nature,  but  being  an  anaerobe,  it  will  not  grow  on  open 
and  exposed  wounds  or  where  there  is  a  free  circulation, 
preferring  deep  punctured  wounds,  such  as  those  produced 
by  the  prong  of  a  pickstaff,  the  point  of  a  shaft  or  hook,  etc. 
It  is  also  seen  in  cases  of  difficult  parturition,  with  injuries 
to  the  vagina  or  vulva. 

Owing  to  the  bacillus  evolving  gas  during  its  growth, 
crepitus  similar  to  that  seen  in  quarter-evil  is  a  common 
symptom. 

The  disease  runs  a  very  acute  course,  and  unless  strong 
measures  are  adopted  very  early,  little  hope  of  recovery 
can  be  entertained. 

A  protective  serum,  having  a  certain  curative  value  if 
u.sed  early,  is  made  by  exposing  the  jugular  vein  of  a  horse, 
and  injecting  increasingly  large  doses  of  virulent  material. 
In  this  way  the  animal  becomes  immune  to  the  pathogenic 
action  of  the  bacillus. 

He  is  then  bled,  and  the  serum  collected  has  prophylactic 
and  curative  qualities  by  reason  of  its  power  of  stimulating 
the  phagocytes,  agglutinins,  and  antitoxins. 

Locally  the  wound  should  be  attended  to.  Free  use  of 
the  knife  should  be  made,  to  allow  all  the  air  possible  to 


130  CLINICAL  BACTERIOLOGY  AND 

enter   the    diseased   tissues,   and,  where  possible,   oxygen 
should  be  injected. 

The  cavities  should  be  irrigated  with  a  solution  of 
potassium  permanganate  or  peroxide  of  hydrogen,  and 
the  circulation  should  be  maintained  by  repeated  doses 
of  strychnine  and  caffeine  hypodermically,  and  orally 
ammonia. 

Blackquarter  (Symptomatic  Anthrax). 

In  some  countries  and  in  certain  counties  this  disease 
affects  a  large  percentage  of  young  cattle;  but,  owing 
to  the  extensive  application  of  vaccine-therapy,  it  is  not 
so  prevalent  as  it  used  to  be  in  pre-vaccination  days. 
Curative  vaccine-therapy  must  ever  occupy  a  secondary 
place  so  far  as  blackquarter  is  concerned.  The  pre- 
monitory symptoms  of  the  disease  are  so  insignificant, 
and  those  that  are  patent  are  of  so  short  duration, 
that  the  immunizer  is  unable  to  use  his  vaccine  until 
the  disease  has  become  established;  and  when  diag- 
nostic lesions  are  in  evidence,  curative  measures,  in  the 
writer's  experience,  are  useless.  As  a  prophylactic,  black- 
quarter  vaccination  has  long  passed  the  experimental  stage, 
and  as  such  its  position  is  now  assured. 

The  bacillus  of  quarter -evil,  technically  known  as  the 
sarcophysematous  hovis  bacillus,  gains  entrance  to  the 
system  through  some  form  of  wound. 

It  is  motile,  anaerobic,  easily  stained,  3  to  6  /j,  long,  and 
usually  shows  at  the  end  a  spore,  giving  the  bacillus  the 
appearance  of  a  club,  resembling  the  bacillus  of  tetanus. 
Unlike  bacilli  in  general,  it  is  Gram-positive ;  but,  like  the 
bacilli  of  malignant  oedema,  during  growth  it  evolves  gas. 

One  attack,  if  the  animal  survives,  confers  immunity  for  a 
long  period,  and  it  is  believed  a  foetus  in  utero  has  complete 
immunity  conferred  upon  it  if  the  mother  contracts  the 
disease  and  lives. 

Several  methods  have  been  adopted  to  produce  satis- 
factory  vaccines,    but    the    two    most    commonly   used   in 


VACCINE-THERAPY  131 

practice  are  Kitt's  single  vaccine  and  Arloing's  double 
vaccine. 

Kitt's  single  vaccine  method  consists  of  attenuating  the 
virus  by  moist  heat  at  a  temperature  of  100°  C.  for  six  or 
seven  hours.  The  affected  muscles  should  be  carefully  dis- 
sected out  and  dried  in  a  dry-air  sterilizer  or  oven  at  a 
temperature  of  35°  C,  after  which  it  should  be  powdered 
and  mixed  with  water  to  form  a  paste.  This  paste  is  now 
spread  on  glass  plates,  and  put  in  a  thermostat  and  subjected 
to  the  temperature  already  stated  above.  It  should  then 
be  dried,  and  the  dose  is  1  decigramme,  mixed  with  sterile 
water  and  a  small  percentage  of  lysol  or  carbolic  acid. 

The  great  advantage  of  this  vaccine  to  the  busy  prac- 
titioner is  that  one  injection  only  is  required;  but  the 
degree  of  immunity  conferred  is  not  so  great  or  so  lasting 
as  that  obtained  in  using  the  double  vaccine.  We  find  in 
practice,  for  economical  reasons,  clients  do  not  like  the 
double  vaccine,  and  if  the  disease  is  not  developing  beyond 
its  usual  sporadic  manner,  the  single  vaccine  answers  quite 
well.  In  some  districts  where  blackquarter  obtains  serious 
proportions,  however,  the  double  vaccine  should  un- 
doubtedly be  used. 

When  vaccination  as  a  prophylactic  was  first  introduced 
into  this  country,  the  writer  had  some  unfortunate  ex- 
periences where  the  subcutaneous  tissue  over  the  shoulder 
was  the  site  chosen  for  injection.  In  several  instances  the 
shoulder  and  leg  swelled  to  alarming  proportions,  and  one 
animal  died,  although  strict  asepsis  was  observed. 

Since  then  we  have  always  used  the  tail  as  the  seat  of 
inoculation.  Here  the  cellular  tissue  is  denser,  absorption 
of  the  vaccine  is  more  gradual,  and  the  danger  to  life  is  nil. 
Against  this,  however,  the  degree  of  immunity  may  not  be 
so  great,  and  in  consequence  a  more  virulent  vaccine  may 
be  used;  and  in  making  our  own  vaccines  now,  we  make 
allowance  for  this  fact  by  subjecting  them  to  a  lower 
temperature. 


132  CLINICAL  BACTERIOLOGY  AND 

Arloing's  Double  Vaccine  Method. 

Here  two  vaccines  of  varying  degrees  of  virulence  are 
used,  the  first  being  given  to  prepare  the  system  for  the 
second  and  more  virulent  dose. 

They  are  prepared  as  follows :  The  diseased  muscle  is 
dried  in  the  sterilizer  (dry-air)  at  a  temperature  of  35°  C, 
powdered,  mixed  with  water  to  form  a  paste,  and  spread 
on  a  glass  plate,  exactly  the  same  as  in  Kitt's  method. 

It  is  then  placed  in  a  thermostat  for  six  hours,  and  main- 
tained at  a  temperature  of  103°  C. 

The  dried  powder  is  now  put  in  a  sterile  mortar, 
ground  down  finely,  placed  in  a  sterile  stock  vaccine  bottle, 
with  rubber  cap  or  glass  stopper,  and  labelled  "Vaccine 
No.  1." 

Vaccine  No.  2  should  be  treated  in  exactly  the  same  way, 
only  instead  of  being  subjected  to  a  temperature  of  103°  C, 
a  temperature  of  95°  C.  is  required. 

The  dose  of  each  vaccine  is  1  centigramme. 

No.  2  vaccine  should  be  administered  within  ten  days 
of  No.  1. 

The  vaccine  may  be  dissolved  in  normal  saline  solution 
to  which  is  added  0*5  per  cent,  of  lysol,  and  used  as  a  liquid 
injection;  or  several  layers  of  thread  may  be  plaited 
together  to  form  a  single  cord.  Several  of  these  should  be 
steeped  in  the  vaccine  emulsion,  each  cord  then  constituting 
one  dose. 

When  an  animal  is  already  infected,  the  vaccine  may  not 
be  effectual — in  fact,  during  the  process  of  vaccination  they 
are  liable  to  bruise  themselves,  and  often  thereby  determine 
the  disease.  In  the  writer's  experience,  complete  immunity 
is  not  arrived  at  until  the  lapse  of  three  weeks,  after  which 
period  they  are  usually  safe  for  twelve  months.  To  reduce 
the  risks  of  bruising  during  inoculation  by  the  cord 
method,  we  find,  if  there  are  a  number  of  animals  to 
be  inoculated,  they  can  be  speedily  done  if  placed  in  a 
small  house,  so  that  they  cannot  run  about.  The  operator 
should  catch  the  tail  of  an  animal  so  sardined  between  its 


VACCINE-THERAPY  133 

fellows  that  it  cannot  move,  and,  by  a  firm  thrust  of  the 
needle,  pierce  the  skin,  taking  care  not  to  drive  the  point 
into  the  coccygeal  bone,  in  which  case  the  needle  invariably 
breaks.  If  done  in  this  way,  the  animals  need  not  be 
caught  or  harassed  in  any  way. 

When  dealing  with  farms  predisposed  to  quarter -evil 
and  pedigree  stock,  we  always  advise  calves  over  six  weeks 
of  age  being  vaccinated,  and  as  a  rule  animals  over  three 
years  of  age  are  usually  proof  against  the  disease,  although 
we  have  known  a  case  in  a  milch-cow  of  six  years  old. 

Anthrax. 

The  extreme  virulence  of  the  B.  anthracis  and  the  high 
percentage  of  fatalities  which  follow  its  invasion  offers 
poor  prospects  of  good  results  following  the  application  of 
curative  measures.  We  find  in  practice  some  animals,  and 
certain  species  of  animals,  which  are  more  resistant  to  its 
destructive  effects,  however,  than  others;  and  it  appears 
the  greater  the  tendency  for  the  disease  to  localize  itself, 
the  more  certain  the  prospect  of  a  recovery.  Algerian 
sheep  seem  to  possess  an  inherited  degree  of  immunity. 
In  cattle  the  disease  usually  shows  itself  as  a  generalized 
septicaemia,  with  consequent  rapid  death. 

In  the  horse  the  tendency  is  for  the  disease  to  confine  its 
effects  to  the  glands  of  the  throat,  and  recovery  has  been 
known  to  follow.  When  the  lesions  are  in  the  bowels  or 
lungs,  death  always  follows. 

In  the  pig  a  favourite  seat  for  infection  is  the  glands  of 
the  throat,  and  not  a  few  recover.  One  attack  seems  to 
confer  a  pronounced  degree  of  immunity. 

Prophylactic  vaccines  have  been  made  by  many  workers, 
but  probably  Pasteur's  vaccine  is  the  most  successful  now 
in  use.     He  prepares  two  vaccines  as  follows  : 

Vaccine  Treatment. 

Vaccine  No.  1  consists  of  cultivating  the  bacilli  on  broth 
at  a  temperature  of  42°  C.  for  twenty-four  days. 


134  CLINICAL  BACTERIOLOGY  AND 

By  so  doing  the  virulence  of  the  bacilli  is  so  attenuated 
that  the  vaccine  so  made  kills  mice,  but  not  guinea-pigs. 

Vaccine  No.  2  is  prepared  in  the  same  manner  as  No.  1, 
save  that  attenuation  goes  on  for  twelve  days  at  the  same 
temperature. 

This  vaccine  destroys  mice  and  guinea-pigs,  but  not 
rabbits. 

For  a  horse  or  ox  j  c.c.  of  vaccine  No.  1  is  first  injected 
into  an  animal,  and  after  ten  to  twelve  days  the  second 
vaccine  is  injected — also  J  c.c.  About  three  weeks  after 
the  last  injection  the  animal  is  proof  against  the  disease, 
which  immunity  lasts  for  about  twelve  months. 

These  vaccines  are  not  altogether  free  from  danger  in 
practice,  and  it  appears  some  animals  are  less  resistant  to 
the  virulence  of  the  vaccine  than  others.  The  writer,  seven 
years  ago,  vaccinated  a  dairy  of  fifty-three  cows  with 
Pasteur's  vaccine,  and  one  case  nearly  died  after  vaccine 
No.  1.     Vaccine  No.  2  was  not  used  on  her. 

An  intense  painful  swelling  occurred  at  the  seat  of  injec- 
tion; the  animal  was  excessively  lame ;  her  breathing  was 
hurried  and  laboured ;  her  temperature  went  up  to  105°  F. 
Full  doses  of  turpentine,  salicylic  acid  and  alcohol,  with 
hypodermic  injections  of  strychnine  and  caffein,  were 
repeatedly  administered,  and  she  eventually  recovered. 
Since  then  we  have  used  the  tail  as  the  seat  of  inoculation. 

Serum  Treatment. 

In  man  several  antiserums  are  in  use  : 

1.  Mendez  immunized  horses  by  injecting  them  over  a 
long  period,  six  to  twelve  months,  after  which  he  bled 
them,  and  obtained  the  serum  in  the  usual  way.  He  records 
excellent  results  by  giving  doses  of  20  c.c.  of  this  serum. 
In  cattle  and  sheep  affected  with  anthrax  he  has  also 
obtained  recoveries  after  using  his  serum. 

2.  Sclaro,  after  a  long  period  of  experimenting,  was 
able  to  immunize  a  goat  against  the  bacillus,  and  produced 
a  protective  and  curative  serum.     Later  he  treated  asses  in 


PLATE   VIII. 


ANTHRAX    BACILLI.        X  1000. 
(Jowett's  "Blood  Serum  Therapy.") 


BACILLUS   TETANI.  X    1500. 

(Hewlett's  "  Bacteriology.") 


To  face  page  134. 


VACCINE-THERAPY  135 

a  similar  manner,  and  obtained  a  serum  which  he  found 
very  efficacious.  He  ascertained,  if  he  injected  2  c.c.  of 
this  serum  into  a  rabbit  along  with  1  c.c.  of  a  fresh  virulent 
broth  culture  of  the  bacillus,  the  animal  did  not  die.  In 
man  he  injected  30  to  40  c.c.  into  the  flank  in  several 
places,  and  noted  a  rise  in  temperature,  followed  by 
recovery.  He  considers  the  serum  is  antibacterial,  and 
stimulates  the  leucocytes. 

3.  Legge  treated  sixty-seven  cases  (British  Medical 
Journal,  1905)  with  this  serum,  and  in  fifty-six  the  serum 
was  used  alone.  He  considers  the  exact  mode  of  action  of 
the  serum  is  unsolved. 

4.  Deutsch  has  also  prepared  a  serum  which  appears  to 
be  effective  in  cattle.  It  agglutinates  the  bacilli,  and  con- 
tains an  immune  body  capable  of  destroying  the  organisms 
in  the  presence  of  a  suitable  complement. 

5.  Sobernhein  has  made  an  antiserum  which  he  com- 
bines with  a  vaccine  similar  to  Pasteur's  No.  2,  and  used 
the  two  at  the  same  time  with  excellent  results. 

This  method  appears  to  be  less  dangerous  than  Pasteur's, 
and,  more  virulent  doses  being  given,  a  greater  degree  of 
immunity  is  arrived  at. 

To  be  efficacious,  it  would  appear,  very  large  doses  of  the 
serum  must  be  given. 


CHAPTEE  XVI 

BACTERIAL  DISEASES  AFFECTING  THE 
NERVOUS  SYSTEM 


Disease. 

Cause. 

Rabies 

...     Ultra- visible  virus. 

Tetanus 

Bacillus  tetani. 

Rabies. 

Fortunately  rabies  at  present  is  an  exotic  disease  in  the 
United  Kingdom,  but  on  the  Continent,  in  India,  America, 
and  Africa  it  is  still  very  rampant. 

It  is  a  specific  infective  disease  due  to  a  filterable  virus, 
affecting-  all  the  domesticated  animals  and  several  of  those 
of  the  wild  species.  The  virus  in  its  most  virulent  form  is 
found  in  the  brain,  the  salivary  glands,  and  the  secretions 
of  the  affected  animal.  The  virus  appears  to  pass  along 
the  nerve  fibres,  and  in  this  manner  reach  the  brain  and 
the  spinal  cord,  where  it  attacks  the  sensory  cells  causing 
irritation  and  producing  symptoms  of  great  excitement,  etc. 
This  condition  of  cellular  irritability  is  followed  by  cellular 
depression  and  organic  degeneration,  and  death  sooner  or 
later. 

There  are  certain  diseases  which  simulate  rabies  in  many 
ways  so  far  as  the  behaviour  of  an  affected  animal  is  con- 
cerned, and  it  is  most  important  that  an  accurate  diagnosis 
should  be  made. 

An  animal  having  been  bitten  and  showing  cardinal 
symptoms  of  rabies,  however,  would  suggest  little  or  no 
difficulty  in  arriving  at  an  accurate  diagnosis.  In  some  cases 
136 


VACCINE-THERAPY  137 

the  history  and  symptoms  are  obscure,  and  further  inquiry 
is  essential  in  order  to  arrive  at  a  positive  conclusion. 

On  microscopical  examination  of  sections  of  the  medulla 
and  cerebellum  of  a  rabid  animal,  Negre  found  certain 
bodies.  These  bodies  stain  pink  Avith  Van  Grieson's  stain, 
the  nerve  cells  are  blue,  and  the  red  corpuscles  colourless 
or  faintly  yellow. 

Further,  if  medullary  extract  of  an  animal  suspected  of 
dying  from  rabies  is  injected  into  another  animal,  and  that 
animal  develops  symptoms  of  rabies,  the  results  are  con- 
clusive. 

Pasteur  was  the  first  to  demonstrate  the  fact  that  a 
recovered  animal  could  not  be  artificially  affected  later 
with  even  large  doses  of  the  active  virus,  and  on  this  basis 
he  introduced  an  antirabic  vaccine.  He  took  a  rabbit 
and  infected  it  by  giving  a  subdural  injection  with  fixed 
virus,  after  which  he  carefully  dissected  out  under  proper 
precautions  the  medulla  and  spinal  cord  and  suspended  it 
in  a  long  tube  at  the  bottom  of  which  was  placed  pieces  of 
caustic  potash,  and  subjected  the  whole  to  a  constant 
temperature  of  23°  C.  By  this  process  it  was  found  the 
cord  became  dry  and  also  lost  a  large  proportion  of  its 
virulence.  Thus  it  was  ascertained  if  a  cord  dried  for  five 
days  in  this  manner  would,  if  introduced  into  an  animal, 
produce  rabies  on  the  eighth;  if  dried  for  nine  days  the 
disease  would  make  itself  manifest  on  the  fifteenth  day; 
and  if  dried  for  fourteen  days  the  effects  of  the  virus  would 
be  nil. 

Therefore  a  medulla  and  cord  subjected  to  heat  for 
fourteen  days  would  give  daily  for  the  same  number  of 
days  fourteen  doses  each,  varying  in  virulence. 

Pasteur  injected  into  dogs  the  most  attenuated  dose  first, 
and  by  diminishing  the  attenuation  he  found  the  time 
came  when  a  complete  degree  of  immunity  was  arrived  at. 
So  much  so,  that  a  dog  so  treated,  if  bitten  by  a  rabid  dog, 
was  proof  against  infection.  Moreover,  he  also  proved  if 
up  to  a  given  time  after  an  animal  had  been   bitten  the 


138  CLINICAL  BACTERIOLOGY  AND 

vaccine  was  injected,  development  of  the  disease  would  be 
prevented,  and  that  immunity  would  be  maintained  for  one 
year  at  least. 

In  the  year  1885  Pasteur,  going  forward  with  the  know- 
ledge he  already  possessed  as  the  result  of  his  experiments 
upon  animals,  successfully  vaccinated  a  boy  who  had  been 
bitten  by  a  rabid  dog. 

Since  then  this  mode  of  prophylactic  treatment  and  its 
modifications  has  been  carried  out  on  an  extensive  scale, 
with  excellent  results.  Hogyes  prepared  a  vaccine  by 
triturating  normal  saline  solution  with  virulent  spinal  cords 
of  rabbits,  making  emulsions  of  different  strengths — i.e. : 

1  in  5,000.  1  in  200. 

1  in  2,000.  1  in  100. 

1  in  500.  1  in  10. 

These  he  injected  every  two  hours,  beginning  with  the 
most  attenuated  dose.  He  found  that  a  dog  so  treated 
would  resist  infection  if  bitten  by  a  rabid  dog  or  injected 
artificially  with  the  virus. 

These  experiments  also  demonstrated  an  interesting  fact 
that  it  is  not  the  attenuation  of  the  virus,  as  in  Pasteur's 
vaccine,  but  a  reduction  of  the  amount  which  is  important. 

Tetanus. 

Tetanus  is  a  specific  infective  disease  due  to  the  action 
of  the  bacillus  of  tetanus  and  its  toxins.  The  bacilli  are 
found  most  commonly  in  garden  soil,  horse  manure,  and 
road  earth,  and  there  must  be  an  injury  to  the  skin  or 
mucous  membrane  before  an  animal  can  become  affected. 

The  intestines  of  the  horse  have  been  found  to  harbour 
the  bacilli  without  producing  a  pathogenic  condition. 

The  bacilli  located  in  a  wound  multiply,  but  seldom  leave 
the  focus  of  infection ;  they  are,  therefore,  rarely  found  in 
the  blood-stream  or  vital  organs.  During  the  process  of 
their  multiplication  they  manufacture  a  specific  toxin,  which 
is  taken  up  by  the  circulation  and  diffused  over  the  entire 


VACCINE-THERAPY  139 

body.  Some  of  the  poison  is  also  carried  along  the  nerve 
fibres.  In  this  manner  also  the  motor  and  sensory  nerve 
cells  become  severely  charged  with  the  poison,  producing 
the  characteristic  symptoms.  This  action  of  the  poison  is 
a  very  difficult  one  to  explain,  but  probably  Ehrlich's  side- 
chain  theory  supplies  the  best  material  that  our  knowledge 
of  the  subject,  as  we  understand  it,  can  give.  Accordingly, 
each  toxin  molecule  is  composed  of  one  non-toxic  (hapto- 
phore)  and  one  toxic  (toxophore)  atom  group.  The 
protoplasm  of  the  motor  nerve  cell  is  composed  of  a  vital 
nucleus  and  numerous  side-chains,  or  receptors,  of  which 
many  possess  a  special  affinity  for  the  haptophore  atom 
groups  of  the  toxin  molecule.  When  such  molecules  of  the 
tetanus  toxin  reach  the  nerve  cells,  they  are  anchored  by 
the  aid  of  the  haptophore  groups  to  the  corresponding 
receptors  of  the  cell  protoplasm,  whereupon  the  toxophore 
group  attack  the  nucleus  of  the  cell.  The  opposed  affinity 
between  tetanus  toxin  and  nerve  substance  is  demonstrated 
by  the  experimental  observation,  where  it  is  shown  that 
a  mixture  of  toxin  and  brain  substance  is  non-toxic  for  a 
guinea-pig  ( Wassermann) . 

When  the  symptoms  of  tetanus  come  on  slowly,  and  the 
disease  itself  appears  to  be  running  an  insidious  course, 
recovery  usually  follows.  On  the  other  hand,  if  the  disease 
is  very  acute  and  the  onset  rapid,  one's  prognosis  must  be 
unfavourable. 

In  the  latter  condition  it  is  probable  the  toxin  has  united 
with  the  cells,  causing  vital  destruction;  while  in  the 
former  it  may  only  have  fixed  itself  to  the  free  side-chains, 
or  receptors,  or  those  that  are  approaching  freedom,  and 
thereby  neutralizing  the  toxic  effects  before  cell  injury  can 
take  place.  It  must  obviously  follow  when  one  remembers 
how  potent  the  toxin  of  tetanus  is ;  to  obtain  any  curative 
results  at  all,  the  poison  must  be  neutralized  before  it 
begins  to  attack  the  cellular  elements.  This  in  practice 
unfortunately  is  not  always  possible,  as  we  have  no 
premonitory  symptoms    to    guide  us,  the  first  indications 


140  CLINICAL  BACTERIOLOGY  AND 

being   a   series   of   established   specific  phenomena  of    the 
presence  of  the  disease  itself. 

On  the  other  hand,  if  a  liberal  liberation  of  receptors 
rapidly  follows,  one  would  expect  that  Nature's  efforts  to 
neutralize  the  toxin  would  take  place. 

In  1890  Behring  immunized  rabbits  by  inoculating  them 
first  with  0'3  c.c.  of  virulent  culture  (filtrated),  and  then  in- 
jecting them  with  3  c.c.  of  a  1  per  cent,  iodine  solution.  Such 
subjects  later  withstood  the  effects  of  a  10  c.c.  injection  of 
virulent  culture,  while  0*5  c.c.  of  the  same  culture  killed 
control  rabbits.  Moreover,  they  also  survived  twenty  times 
the  fatal  dose  of  toxin,  while  0*2  c.c.  of  their  blood-serum 
protected  them  against  a  virulent  infection  given  twenty- 
four  hours  later.  These  experiments  laid  the  founda- 
tion upon  which  serum-therapy  as  a  prophylactic  and 
curative  method  was  built. 

Behring  adopted  the  following  method  in  making  his 
vaccine  serum :  He  took  a  virulent  broth  culture  of  the 
bacillus,  and  in  80  c.c.  he  mixed  0-25  per  cent,  trichloride 
of  iodine;  60  c.c,  0*175  per  cent,  iodine;  40  c.c,  0*125  per 
cent. ;  while  he  retained  another  20  c.c.  which  he  used 
without  mixing  at  all.  He  then  injected  subcutaneously 
into  the  horse  the  most  attenuated  culture  first,  and  pro- 
ceeded with  the  others  at  intervals  of  eight  days. 

Finally  he  injected  the  pure  culture,  0*5  c.c,  doubling  the 
dose  every  five  days. 

After  the  lapse  of  a  given  time  the  animal  was  bled,  and 
its  serum  obtained  in  the  usual  way. 

An  animal  injected  with  an  immune  serum  lasts  from 
three  to  four  weeks.  The  serum  may  be  administered 
subcutaneously,  intravenously,  or  intracranially,  and  as  such 
possesses  antitoxic  properties. 

For  reasons  already  explained,  the  action  of  antitetanic 
serum  as  a  curative  agent  has  considerable  limitations,  and 
in  veterinary  practice,  at  least,  its  usefulness  is  mainly 
prophylactic. 

Where  we  have  contused  wounds  or  injuries,  caused  by 


VACCINE-THERAPY  141 

rusty  rails,  wire  fencings,  agricultural  tools  and  implements, 
etc.,  a  dose  of  antitetanic  serum  injected  will  produce 
complete  protection  for  three  or  four  weeks. 

In  addition,  such  wounds  should  be  cauterized  and  even 
scarified,  and,  above  all,  severely  dressed  with  very  strong 
solutions  of  tincture  of  iodine.  If  the  infective  wound  is 
low  down  the  leg  (below  the  knee),  in  such  cases  we  believe 
we  have  had  benefit  by  performing  neurectomy.  The 
application  of  Bier's  treatment,  recommended  by  some  to 
check  absorption,  we  consider  detrimental  to  the  patient. 
This  procedure  causes  an  increase  of  pain  to  the  patient 
and  aggravates  the  tetanic  symptoms. 

Where  lockjaw  is  the  result  of  an  amputated  tail, 
we  reamputate,  taking  off  several  inches,  if  possible,  and 
inject  tincture  of  iodine  at  the  root  of  the  tail. 

The  intratraehial  injections  of  lugol  solution  we  have 
found  not  only  to  mitigate  the  severity  of  the  attack  in 
some  cases,  but  in  those  cases  which  do  recover,  the  duration 
of  convalescence  is  considerably  shortened. 

The  subcutaneous  injections  of  mag.  sulph.  in  solution 
in  our  hands  have  proved  of  no  curative  value,  and  the 
same  may  be  said  of  the  rectal  injections  of  liquor  ferri 
perchlor. 


CHAPTER  XVII 
DISEASES  OF  THE  RESPIRATORY  ORGANS 


Pneumonia 

Strangles     ... 
Purpura  hemorrhagica 

Influenza    ... 

Glanders     ... 
Distemper  ... 


Actinomycosis 
Phthisis 


Disease.  Cause. 

Bronchitis Pneumococcus,     streptococcus,     Micro- 
coccus catarrhalis. 

Pneumococcus,  streptococcus,  staphylo- 
coccus, B.  coli,  cocco  bacillus. 

Streptococcus  and  others. 

Staphylococcus,  streptococcus,  diplococci, 
and  others. 

Primary  :  ultra- visible.  Secondary  strep- 
tococcus, staphylococcus. 

B.  mallei. 

Primary:  ultra- visible  or  Bronchosep- 
ticus.  Secondary  streptococci,  staphy- 
lococcus. 

Streptothrix  actinomyces. 

B.  tuberculosis.  Secondary  infection 
may  include — Streptococci,  staphylo- 
cocci, pneumococcus,  Micrococcus 
catarrhalis,  M.  tetragenus,  B.  coli. 

Bronchitis. 

It  may  be  taken  for  granted,  so  commonly  is  the  fact  seen 
in  practice,  that  bacterial  invasion  of  the  respiratory  tract, 
from  the  anterior  nares  to  the  pulmonary  vesicles,  is  a 
mixed  one ;  indeed,  the  respiratory  mucosas  continually 
harbour  pathogenic  bacteria  whose  disease  -  producing 
qualities  only  become  manifest  through  some  disturbance 
of  Nature's  protective  and  bacteriotropic  forces.  No  sooner 
does  a  specific  organism  gain  a  footing,  and  disease  sets  in, 
than  other  bacteria  have  placed  before  them  increased 
opportunities  for  their  development,  which  opportunities 
they  are  not  slow  to  embrace.  All  atmospheres  are  charged 
142 


VACCINE-THERAPY  143 

with  bacteria,  but  some  more  so  than  others.  The  badly 
ventilated  horse-box,  the  fusty  cow-pen,  or  the  insanitary 
dog-kennel,  pre-eminently  carry  more  than  their  share,  and, 
needless  to  add,  animals  with  pulmonary  disease  inhaling 
such  germ-laden  air  must  be  placed  at  great  disadvantage 
when  the  crisis  of  the  disease  arrives.  In  fact,  we  see  in 
practice,  some  cases  where  specific  invasions  of  the 
respiratory  organs  have  taken  place  under  excellent 
hygienic  conditions  making  good  recoveries,  while  others, 
less  fortunate  by  reason  of  their  unhygienic  surroundings, 
succumb  to  pulmonary  disease  in  spite  of  all  efforts  to  save 
them.  The  first  point,  then,  in  all  respiratory  affections 
the  clinician  must  emphasize  is  to  insist  his  patient  inhaling 
the  purest  possible  air-supply. 

Diseases  of  the  respiratory  apparatus  seem  to  be  particu- 
larly suited  to  the  beneficial  influences  of  vaccine-therapy, 
the  circulatory  supply  being  usually  so  good,  unless,  of 
course,  some  mechanical  or  other  obstructive  lesions  are 
in  existence  to  prevent  the  bactericidal  elements  easily 
reaching  the  source  of  infection. 

Moreover,  the  clinical  phenomena  after  vaccine  injection 
can  usually  be  more  easily  interpreted  here  than  probably 
in  any  other  system  of  organs. 

Perhaps  the  most  common  group  of  exciting  causes 
productive  of  derangement  of  the  respiratory  system  are 
sudden  barometric  changes. 

A  sudden  chilling  of  the  respiratory  mucosee  drives  the 
blood  and  its  antibodies  temporarily  away  from  the  parts, 
the  previously  latent  bacteria  often  found  located  upon 
the  respiratory  mucosas  become  active,  and  in  the 
process  of  their  growth  toxins  are  formed  which  still  more 
depress  the  body- cells.  Local  cell-proliferation  goes  on, 
leucocytosis  comes  into  evidence,  and  the  whole  area  is 
bathed  in  bacteriotropic  material.  A  regular  warfare  has 
now  become  established,  and  upon  the  power  of  the  tissue- 
elements  to  overthrow  the  bacteria  depends  whether  the 
animal  lives  or  dies. 


144  CLINICAL  BACTERIOLOGY  AND 

When  one  is  dealing  with  a  mixed  infection,  it  is  obvious, 
if  vaccine-therapy  is  to  serve  its  intended  purpose,  a 
vaccine  of  all  the  causative  bacteria  must  be  obtained.  In 
practice  it  is  difficult  to  estimate  to  each  bacterium  the 
degree  of  pathogenicity  which  that  particular  bacterium 
has  the  power  of  exercising,  and  it  is  probable  that  in  each 
individual  case  the  degree  varies  considerably  in  the  same 
species    of    bacteria   at    different   times   and  in    the   same 


To  ascertain  the  nature  of  infection,  one  must  collect  the 
pathological  material  at  one's  disposal.  In  animals  we 
obtain  such  from  the  nasal  discharges,  and  the  materis 
morbi  should  be  carefully  collected,  to  prevent  outside  con- 
tamination, as  already  described. 

If  the  patient  has  had  a  good  previous  health  record,  we 
begin  by  giving  the  following  doses,  assuming,  of  course, 
all  these  bacteria  are  present : 

Staphylococci 500,000,000 

Streptococci      100,000,000 

Diplococci         500,000,000 

Micrococcus  catarrhalis  . . .  250,000,000 

The  clinical  phenomena  should  be  carefully  noted. 

Some  twelve  hours  after  injection  the  temperature  will 
usually  rise  one  or  two  degrees,  the  pulse-beat  will  be 
increased  in  number  10  to  20  per  minute,  and  the  respiration 
increases  5  to  15  per  minute. 

The  cough  is  usually  more  irritable  and  the  catarrhal 
discharge  increased  in  quantity,  but  more  tenacious  in 
consistence.  On  the  whole,  there  is  a  proportionate  general 
depression.  To  sum  up,  these  phenomena  constitute  the 
negative  phase,  which  are  salutary  if  the  vaccine  has 
responded  in  the  manner  one  desires,  and  they  are  to  be 
expected  if  one  anticipates  good  results  to  follow.  If  no 
such  characteristics  are  noticed,  the  dose  is  too  small  or 
there  is  some  error  in  the  technique,  and  a  close  investiga- 
tion is  called  for. 


PLATE   IX. 


•  <    \ 


MICROCOCCUS   CATARRHALIS— HUMAN    SPUTUM. 

(Emery's  "Clinical  Bacteriology.") 


PNEUMOCOCCI. 
(Itose  and  Carless'  "  Manual  of  Surgery.") 


To  face  page  144. 


VACCINE-THERAPY  145 

If  it  is  due  to  the  smallness  of  the  dose,  an  increased 
dose  should  be  given  straight  away,  amounting  to  from  one- 
half  to  double  the  strength  of  the  original  dose. 

Following  upon  the  negative  phase,  a  steady  improve- 
ment should  be  noted.  The  temperature  falls ;  the  respira- 
tion becomes  slower  and  deeper;  the  pulse -beat  lessens, 
etc.  This  is  the  positive  phase,  which  should  go  on  for 
two,  three,  or  five  days. 

As  soon  as  the  immunizer  notes  a  cessation  of  the  im- 
proving symptoms,  another  dose  of  vaccine  should  be 
given. 

It  is  our  experience  in  pulmonary  conditions  that  vaccine- 
therapy  gives  a  bigger  percentage  of  recoveries,  the 
disease  cycle  is  cut  shorter,  and,  when  recovery  is  an 
accomplished  fact,  the  after-sequela  are  either  nil  or  very 
slight — in  other  words,  permanent  organic  disease  is  not  so 
likely  to  be  left  behind. 

In  severe  cases  several  injections  may  have  to  be  given, 
and  where  progress  is  not  satisfactory,  the  vaccine  should 
be  combined  with  a  polyvalent  antistreptococcic  serum, 
beginning  with  doses  of  20  c.c,  and  increasing  daily 
according  to  results. 

Of  course,  in  the  adaptation  of  vaccine-therapy — and  this 
remark  applies  not  only  to  pulmonary  diseases  in  particular, 
but  to  all  other  bacterial  infection  in  general — orthodox 
treatment  must  not  be  altogether  ignored.  In  addition, 
therefore,  it  is  an  imperative  necessity  to  place  a  lung- 
diseased  patient  in  the  best  hygienic  surroundings,  for 
reasons  already  explained.  One  must  also,  if  it  is  considered 
advisable  and  the  case  warrants  such  a  course,  apply 
counter-irritants  to  the  chest,  administer  circulatory  seda- 
tives or  stimulants,  according  to  the  condition  of  the 
patient — expectorants  if  the  cough  is  troublesome  or  the 
discharge  is  tardy,  antizymotic  febrifuges  if  the  tem- 
perature requires  pulling  down,  and  antiseptic  inhalations 
when  advisable. 

Nevertheless,  when  all  these  so-called  curative  remedies 

10 


146  CLINICAL  BACTERIOLOGY  AND 

are  applied — in  fact,  if  they  were  all  collected  and  totalled 
into  one  and  placed  against  the  antidotal  and  protective 
elements  of  Nature — one  wonders  what  the  ratio  would 
work  out  at  as  regards  their  respective  curative  values. 
We  would  be  inclined  to  place  the  therapeutic  value  very 
low  indeed,  and  this  fact  must  never  be  overlooked  in  what- 
ever line  of  therapeutic  action  we  like  to  take  up. 

In  short,  how  far  can  antizymotics  be  pushed  in  the 
supposed  attempt  to  destroy  bacteria  circulating  in  the 
blood  or  incubating  on  the  respiratory  mucosae,  and 
leave  the  leucocytes  active  and  intact  ?  Or  to  .what 
extent  does  the  inhalation  of  antiseptic  vapours  limit 
the  destructive  processes  of  bacteria  located  in  the  respi- 
ratory mucosae,  or  retard  their  rapid  development  ?  One 
must  confess  those  orthodox  means  are  futile — nay,  more, 
if  they  are  pushed  too  far,  they  may  become  elements  of 
positive  harm,  hastening  on  the  retrogressive  changes  which 
lead  to  dissolution. 

We  must,  then,  fall  back  upon  the  protective  antibodies 
of  the  blood  to  supply  our  antidotal  remedies,  and  the 
more  we  foster  and  stimulate  them,  the  more  likely  we  are 
to  effect  recoveries. 

Pneumonia. 

Every  practitioner  knows  the  orthodox  treatment  of  a 
severe  case  of  pneumonia  is  far  from  satisfactory. 

There  are  no  specific  drugs  upon  whose  value  we  should 
like  to  greatly  rely,  and  after  attending  to  the  patient's 
comfort,  sustaining  him  with  nutrients  to  give  him 
strength  to  face  the  crisis,  and  placing  him  in  good  hygienic 
surroundings,  we  are  practically  at  the  end  of  our  resources, 
and  have  to  fall  back  upon  the  vis  medicatrix  nature. 

In  practice  we  find  pneumonia  occurring  as  a  complica- 
tion to  such  bacterial  diseases  as  strangles  and  influenza, 
or  it  may  be  seen  as  an  independent  and  primary  condition. 
Again,  we  may  find  a  single  isolated  case  in  the  stud,  or 
several  may  become  affected ;  but  whatever  type  the  disease 


VACCINE-THERAPY  147 

may  be,  it  is  clearly  one  for  careful  bacteriological  in- 
vestigation at  the  outset. 

Lignieres  in  1897  discovered  a  bacillus  of  the  pasteurella 
group  which  he  called  the  "  coccobacillus,"  and  which  he 
considered  to  be  the  causative  organism  of  catarrh,  bron- 
chitis, pneumonia,  strangles,  purpura  hemorrhagica,  and 
by  its  action  upon  the  mucosa,  and  through  it  the  system, 
depressing  the  tissues  locally,  and  the  constitution  generally , 
that  the  more  pathogenic  bacteria  gain  a  footing,  with 
serious  functional  and  organic  consequences,  after  which 
the  coccobacilli  are  supposed  to  drop  into  insignificance. 

Be  this  as  it  may,  we  know  for  a  fact  that  in  typical 
pneumonias  mixed  infections  are  the  rule. 

As  one  is  generally  called  in  to  see  the  disease  in  its 
acute  form,  it  is  obvious  there  is  little  time  for  delay,  and  a 
stock  mixed  vaccine,  preferably  polyvalent,  should  be 
injected  at  the  outset.  This  will  give  the  immunizer  time 
to  make  an  autogenous  vaccine,  and  this  rule  we  rigidby 
carry  out  in  practice. 

The  phenomena  after  injection  are  practically  the  same 
as  in  bronchitis,  and  in  point  of  fact  the  two  diseases  run  a 
close  parallel,  so  much  so  that  a  bronchitis  at  any  moment 
may  merge  into  a  pneumonia,  constituting  broncho-pneu- 
monia. We  strongly  believe  the  verminous  pneumonias 
one  sees  in  cattle  and  sheep  derive  their  fatal  consequences, 
not  so  much  from  pulmonary  inflammations  due  to  the 
irritation  of  these  parasites  as  to  the  invasion  of  pathogenic 
bacteria  at  a  later  stage  of  the  disease,  and  in  such  cases 
as  these  we  have  isolated  staphylococci,  streptococci,  and 
diplococci. 

When  a  pneumonia  is  running  a  very  acute  course,  with 
a  high  temperature,  it  is  strongly  advisable  to  begin  with 
a  very  small  initial  dose ;  and  the  reason  is  obvious :  after 
a  vaccine  injection  the  opsonic  cycle  drops  (negative 
phase),  and  the  temperature  rises;  conversely,  with  a  rise 
of  the  opsonic  cycle  (positive  phase)  the  temperature 
drops ;  but  if  the  temperature  is  naturally  high  during  the 


148  CLINICAL  BACTERIOLOGY  AND 

negative  phase,  it  will  go  higher  at  this  period,  and  it  is 
then  the  danger  of  a  dose  too  large  for  Nature  to  assimilate, 
so  to  speak,  comes  in.  This  is  a  principle  applicable  to  all 
acute  diseases,  as  has  already  been  pointed  out,  so  far  as 
vaccine-therapy  goes. 

Strangles. 

This  disease  is  very  prevalent  in  young  horses  at  certain 
seasons  of  the  year,  and  in  some  years  more  so  than  in 
others.  It  is  a  specific  infective  fever,  and  when  running 
a  simple  course  it  affects  the  upper  air-passages  and  in- 
vades the  neighbouring  local  lymphatic  system. 

The  streptococcus  which  is  responsible  for  this  condition 
is  found  in  the  nasal  organs,  and  possibly  exists  as  a  non- 
pathogenic bacterium  in  many  instances.  The  disease  may 
run  a  very  mild  course,  and  pass  off  only  as  a  simple 
catarrh.  This  may  be  due  to  the  low  virulence  of  the 
bacteria  themselves,  or  the  increased  resisting  powers  of  the 
antibodies  of  the  blood  itself,  or  both. 

In  an  ordinary  simple  case  of  strangles  special  treatment 
is  not  called  for,  and  all  that  is  necessary  is  to  keep  the 
patient  from  contracting  chills.  Complications  are,  how- 
ever, liable  to  supervene,  in  which  case  they  must  be 
treated  upon  their  merits. 

If  the  disease  runs  an  irregular  course,  the  immunizer 
can  do  much  to  fortify  his  patient  and  prevent  septicaemic 
or  metastatic  lesions  developing  by  the  injection  of  an 
autogenous  vaccine. 

Here,  too,  if  the  disease  is  one  of  considerable  standing, 
a  mixed  infection  has  to  be  looked  for ;  in  which  case  the 
causative  bacteria  must  be  identified,  and  suitable  vaccines 
made. 

When  Ave  have  under  treatment  a  bad  case,  which 
does  not  seem  to  respond  to  vaccine  treatment,  we  make 
it  a  practice  of  combining  the  vaccine  with  antistrepto- 
coccic serum. 


PLATE  X. 


STREPTOCOCCUS     OF    STRANGLES 

ITS  FROM  A  SUBMAXILLARY  , 


GLANDERS    BACILLI'S.  X   1000. 

(Hewlett's  "Bacteriology.") 


lo  face  page  14n 


VACCINE-THERAPY  149 

Antistreptococcic  Serum. 

It  has  been  proved  that  after  injecting  an  emulsion  of 
devitalized  streptococci  into  the  circulation  of  a  horse,  and 
following*  this  up  by  small  and  gradually  increasing  doses 
of  living  bacteria,  the  time  arrives  when  that  animal's 
serum  has  acquired  high  antimicrobic  powers.  Marmorek 
was  the  first  to  demonstrate  this  fact ;  and  he  also  showed,  if 
the  antistreptococcic  serum  thus  obtained  is  injected  into  an 
animal,  and  that  animal  is  subjected  to  the  action  of  viru- 
lent streptococci,  it  will  prove  itself  to  be  either  completely 
resistant,  or,  if  the  disease  should  develop,  it  will  show  itself 
in  an  exceedingly  mild  form. 

It  is  most  important  to  remember  that  there  are  many 
strains  of  streptococci,  and  each  may  be  equally  virulent ; 
and  one  can  demonstrate  this  from  their  appearances  easily 
under  the  microscope.  Take,  for  example,  the  plump,  well- 
nourished,  globular-looking  types  one  sees  in  a  severe 
attack  of  strangles,  and  compare  them  with  the  shrivelled- 
up,  small,  almost  oval-shaped,  short,  compressed  chains  as 
seen  in  those  cases  of  bovine  endometritis.  In  making, 
therefore,  a  satisfactory  serum,  a  polyvalent  serum  is  to  be 
preferred  to  a  monovalent  one.  It  is  the  experience  of  the 
writer,  in  the  cases  where  the  virulence  of  the  bacteria  is 
very  great,  that  a  combination  of  an  autogenous  vaccine 
and  a  stock  serum  (polyvalent)  gives  much  better  results 
than  if  either  is  used  singly. 

In  fact,  so  strongly  do  we  hold  this  view  now,  that,  given 
a  serious  case  with  septicasmic  tendencies,  we  never  think 
of  failing  to  combine  the  serum  with  the  vaccine. 

In  some  strangles  outbreaks  the  disease  takes  on  a  very 
virulent  form,  and,  in  addition  to  manifesting  local  lesions, 
extension  takes  place,  probably  through  the  lymphatic 
system,  to  the  mesenteric  or  mediastinal  glands. 

Or  some  vital  organ,  such  as  the  liver,  kidneys,  etc., 
may  become  involved.  When  the  blood-stream  itself  has 
become  invaded  by  the  bacteria,  a  typical  septicaemia  has 


150  CLINICAL  BACTERIOLOGY  AND 

taken  place,  and  the  consequences  are  then  generally 
disastrous. 

In  those  cases  where  pronounced  local  abscess  forma- 
tions are  the  order  of  things,  it  is  imperative  the  pus 
cavities  should  be  evacuated  as  soon  as  possible.  By 
so  doing  internal  pressure  is  relieved  from  the  abscess 
wall,  thereby  allowing  the  bactericidal  elements,  by  a 
process  of  osmosis,  to  reach  the  infective  centres.  It  also 
tends  to  prevent  the  risk  of  metastatic  invasions,  a  condition 
always  to  be  feared  and  warded  against  in  strangles. 

We  usually  begin  by  giving  an  autogenous  vaccine 
of  streptococci,  composed  of  250,000,000  as  a  dose,  and  if 
the  case  is  sufficiently  bad  we  add  20  c.c.  of  antistrepto- 
coccic serum. 

The  serum  we  repeat  the  next  day  (30  c.c),  noting  the 
temperature,  pulse,  etc.  On  the  third  day,  if  necessary, 
we  give  40  c.c,  and  if  the  case  is  progressing  we  leave  off 
doing  anything  further  until  the  fifth  day,  when  we  repeat 
the  vaccine,  giving  500,000,000.  If  the  negative  phase  is 
pronounced,  it  is  better  to  wait  and  note  progress  for 
three  to  five  days  before  giving  another  500,000,000  to 
1,000,000,000  for  a  dose.  If  the  negative  phase  is  only 
indifferent,  do  not  wait  so  long,  but  within  twenty-four 
hours  give  500,000,000  to  750,000,000,  and  watch  the 
result.  In  obstinate  cases  one  may  have  to  follow  up 
treatment  for  a  considerable  period,  increasing  the  doses 
as  one  goes  along,  and  watching  the  phenomena. 

The  dose  of  serum,  and  the  number  of  times  it  should  be 
administered,  must  be  items  left  largely  to  the  individual 
immunizer  upon  which  to  exercise  his  judgment. 

Purpura  Hemorrhagica. 

It  is  probable  this  disease  in  equines  is  never  seen  as 
a  primary  condition,  but  is  met  with  in  practice  as  secon- 
dary to  such  specific  debilitating  disorders  as  influenza, 
strangles,  etc. 

That  the  phenomena  produced  are  due  either  to  a  direct 


VACCINE-THERAPY  151 

or  an  indirect  effect  of  one  or  more  species  of  bacteria  or 
their  products  no  one  can  deny,  but  that  the  bacteriology 
of  the  disease  leaves  a  great  deal  to  be  desired  we  must 
all  admit. 

In  cases  of  this  kind  a  variety  of  micro-organisms  have 
been  isolated  from  time  to  time,  including  diplococci, 
staphylococci,  streptococci,  and  bacilli,  and,  judging  from 
our  present  bacteriological  knowledge,  it  is  difficult  to 
ascribe  to  which  microbe  the  production  of  the  specific 
blood-lesions  belongs. 

When  one  is  dealing  with  a  simple  catarrhal  fever,  an 
attack  of  influenza  (so  called),  or  an  outbreak  of  strangles, 
in  a  stud  of  horses,  the  disease  generally  attacks  one 
animal  first,  the  others  slowly  or  suddenly  developing  the 
affection.  As  the  disease  goes  through  the  stable,  one  can  in- 
variably isolate,  microscopically  and  biologically,  the  specific 
microbe,  or,  in  the  case  of  mixed  infections,  microbes  from 
all  the  patients.  As  a  sequel  to  the  outbreak  one  animal 
may  develop  purpura  heemorrhagica,  for  in  practice  we 
seldom  see  more  than  one  case,  the  disease  being  very 
sporadic,  and  all  the  others  may  make  a  good  recovery. 

Why  should  this  one  animal  be  singled  out  for  this  com- 
plication ?  He  has  been  under  the  same  dietetic,  hygienic, 
and  therapeutic  management,  and  his  constitutional  health 
and  age  before  the  illness  began  Was  as  good  as  the  others 
which  recovered.  Nay,  more ;  for  all  that  we  know  the 
clinical  phenomena  suggested  that  the  specific  infection 
from  which  he  was  suffering  was  identical  to  the  others 
which  recovered  without  this  complication. 

We  are  now  face  to  face  with  one  of  two  facts  :  either 
this  animal  has  had,  firstly,  a  hypervirulent  dose,  or,  secondly, 
his  protective  forces  are  so  defective  that,  not  only  is  he 
unable  to  resist  the  invasion,  but  the  specific  invading 
bacteria  find  in  his  economy  the  very  essentials  in  the  most 
suitable  proportion  to  suit  their  requirements. 

As  regards  the  first  hypothesis,  it  is  scarcely  tenable  ; 
for  why  should    this    animal   be   singled  out   for    a   more 


152  CLINICAL  BACTERIOLOGY  AND 

virulent  dose  of  infection  than  the  others,  considering  their 
surroundings  are  identical  and  the  infection  apparently  the 
same  ?  Clearly,  then,  there  must  be  in  this  animal  some 
defective  condition  which  was  non-existent  in  the  others. 

As  we  have  already  seen,  purpura  makes  itself  manifest 
at  the  tail  end  of  debilitating  specific  diseases.  We  know 
a  regular  war  has  been  going  on  between  the  bactericidal 
forces  of  the  body  on  the  one  hand,  and  the  bacteria  on 
the  other. 

The  serum  of  the  blood,  as  we  have  seen,  provides  these 
forces,  and  usually  after  a  bacterial  attack  the  bacterio- 
tropic  indices  rise  and  the  patient  becomes  more  resistant ; 
but  this  degree  of  specificity  is  only  potent  to  the  bacteria 
present,  and  while  these  immune  bodies  are  being  elaborated 
for  the  purpose' of  protecting  the  system  against  the  active 
pathogenic  bacteria,  a  latent  bacterium  makes  its  presence 
felt  by  taking  on  a  pathogenic  role,  the  antibodies  are  not 
sufficiently  prepared  to  resist  this  fresh  invader,  the 
blood- serum  undergoes  a  retrogressive  change,  the  endo- 
thelial cells  also  become  affected  in  some  way,  and  the 
characteristic  petechial  lesions  and  the  serous  effusions 
soon  follow.  This,  of  course,  is  only  hypothesis,  but  we 
venture  to  think  we  must  look  to  some  alteration  of  the 
serum  for  the  production  of  the  purpuric  lesions.  The 
point  is,  what  is  the  nature  of  the  alteration  and  how  is  it 
brought  about  ? 

For  vaccine- therapy  to  be  of  any  service  in  purpura,  we 
must  be  somewhat  empirical  in  our  methods,  and  we  will 
continue  to  be  so  until  we  know  more  of  the  pathology  of 
this  disease.  In  the  meantime  it  becomes  us  to  utilize  what 
knowledge  we  possess  to  protect  our  patient  from  the 
destructive  influences  of  this  fatal  disease. 

Autogenous  vaccines  should  therefore  be  made  from  the 
bacteria  which  have  caused  the  original  condition,  and  they 
should  be  combined  with  serums  if  considered  advisable. 

The  intratracheal  injections  of  Lugal's  solution  must  also 
be  stronglv  recommended  as  an  adjuvant. 


VACCINE-THERAPY  153 

In  fact,  in  days  gone  by,  when  one  used  to  see  more 
cases  of  purpura  than  one  does  now,  LugaPs  solution  was  a 
most  useful  preparation,  and  in  the  writer's  hands  gave 
many  good  results. 

Equine  Influenza. 

The  word  "  influenza,"  as  used  in  veterinary  medicine, 
has  a  very  wide  meaning,  and  as  such,  in  many  cases,  is 
undoubtedly  a  misnomer — so  much  so  that  the  clinician 
recognizes  a  variety  of  diseases  which  may  come  under  the 
same  heading.  This  is  unfortunate,  inasmuch  as  it  admits 
of  laxity,  and  serves  no  useful  purpose,  so  far  as  descriptive 
details  go. 

Certain  it  is  an  endeavour  should  be  made  to  curtail  the 
meaning  of  the  word,  and  at  the  same  time  there  should 
be  drawn  up  a  list  of  cardinal  symptoms  and  lesions  specific 
in  themselves  to  the  disease  in  question.  In  this  way  the 
meaning  of  the  word  itself  would  be  more  confined,  and  the 
information  it  conveyed  more  accurate.  One  cannot  help 
recognizing,  however,  there  are  certain  difficulties  which 
will  remain  until  we  are  in  possession  of  a  more  accurate 
knowledge  regarding  the  nature  of  the  causative  bacteria. 
Some  authorities  hold  the  disease  proper  is  due  to  a  microbe 
of  the  ultra-visible  group ;  others,  that  it  is  caused  by  a 
coccobacillus  which  is  always  present  in  the  primary 
infective  stage,  and  which  is  followed  later  by  a  secondary 
infection,  the  causative  bacterium  in  this  secondary  infec- 
tion in  most  cases  being  a  streptococcus. 

Admitting,  then,  the  bacteriology  of  influenza  is  in  such 
a  nebulous  condition,  one  would  suggest  that  here  at  least 
the  immunizer  has  reached  his  limitations,  and  at  first  sight 
this  would  appear  to  be  so.  On  looking  more  closely  into 
the  facts,  however,  what  do  we  find  in  practice  ? 

Every  practitioner  with  experience  of  influenza  outbreaks 
will  agree  that  the  disease  influenza  itself  is  in  nearly  every 
instance  a  benign  one,  and  that  fatal  consequences  from 
the  primary  and  direct  effects  of  the  disease  are  practically 


154  CLINICAL  BACTERIOLOGY  AND 

nil.  We  must  look,  then,  for  the  sequelae  of  influenza  to 
account  for  the  percentage  of  fatalities;  and  this  is  just 
what  we  see  in  practice. 

If  we  are,  therefore,  in  a  position  to  fortify  the  system 
against  the  causative  bacteria  producing  the  complications, 
obviously  we  shall  succeed  in  reducing  the  death-rate  to 
a  large  degree.  The  more  important  complications,  as  we 
all  know,  are  pneumonia,  purpura,  strangles. 

In  the  majority  of  influenzal  outbreaks  in  a  stud,  one 
animal  only  at  the  outset  sickens,  and  most  probably  this  is 
the  carrier  of  the  infection.  In  the  course  of  one  or 
several  days  others  may  show  premonitory  symptoms. 
This  clearly  gives  to  the  practitioner  a  clue  to  the 
degree  of  infectivity  of  the  virus  on  the  one  hand,  and 
the  natural  resisting  forces  of  his  patients  on  the  other. 
In  addition,  the  first  animal  to  become  affected  affords 
material  for  a  bacteriological  investigation  on  the  part  of 
the  practitioner,  which  the  writer  submits  he  owes  as  a  duty 
to  his  client  and  to  his  own  reputation. 

For  by  so  doing  an  outbreak  not  only  might  be  robbed 
of  its  power  of  producing  monetary  losses  in  the  shape 
of  sacrificed  lives,  but  the  animals  themselves,  if  they 
became  affected,  would  be  restored  to  work  much  sooner  by 
using  up-to-date  prophylactic  and  curative  measures. 

In  these  days  it  is  no  credit  to  us,  as  a  profession,  to  lose 
patients  from  the  complicated  sequels  following  upon  in- 
fluenza, and  it  is  clearly  our  duty  to  ward  against  such  risks. 

How  is  this  to  be  done  ?     By  immunization. 

A  careful  microscopical  examination  should  be  made 
of  the  morbid  material  taken  from  the  nostrils,  if  any; 
the  purulent  discharge  from  an  abscess,  if  present ;  and,  if 
need  be,  a  microscopical  investigation  of  the  patient's  blood 
should  be  carried  out.  In  this  way  a  fairly  accurate  diag- 
nosis of  the  bacteria  present  will  be  arrived  at.  Biological 
investigations  should  now  be  instituted  with  a  view  to 
confirming  and  adding  to  the  microscopical  diagnosis,  and 
also  preparatory  to  the  making  of  the  necessary  vaccines. 


VACCINE-THERAPY  155 

It  may  be  taken  for  granted,  if  one  horse  in  a  stud  or 
stable  is  suffering  from  influenza,  others  are  sure  to  follow, 
and  that,  in  fact,  they  are  invariably  already  incubating  the 
disease. 

Should  the  outbreak  be  a  severe  one,  with  a  tendency  to 
the  development  of  sequelae,  every  animal  should  have,  as 
a  prophylactic,  an  injection  of  a  mixed  vaccine,  preferably 
derived  from  the  cultivated  bacteria  obtained  from  the 
first  animal  to  succumb  to  the  disease. 

Of  course,  it  does  not  follow  every  animal  will  show  the 
same  variety  of  bacteria,  even  in  the  same  stable ;  but  it  may 
be  taken  for  granted  that  a  vaccine  derived  from  a  patient 
suffering  from  the  disease  similar  to  the  infection  to  which 
the  others  have  been  subjected,  and  under  the  same  roof,  is 
more  likely  to  contain  the  identical  or  analogous  bacteria 
than  if  procured  from  a  patient  exotic  to  the  diseased  area 
or  building. 

If  streptococci  are  present,  it  is  an  advantage  to  combine 
antistreptococcal  serum  with  the  vaccine. 

When  an  animal  is  suffering  severely,  and  sequelae  set 
in,  an  autogenous  vaccine  should  be  made  from  the  dis- 
charges, and  the  animal  treated  according  to  its  condition. 
In  the  few  cases  in  which  we  have  taken  the  opsonic 
index  in  influenza  sequelae,  it  has  invariably  been  very 
low  to  streptococci,  and  in  one  case  it  was  only  039. 

Three  days  after  injection  another  estimate  of  the 
opsonic  content  was  made,  and  it  was  found  to  have  risen 
to  1-57. 

So  far  as  influenza  and  strangles  are  concerned,  we  have 
already  seen,  grave  sequelae  may  follow  either,  and  in 
many  cases  with  fatal  results.  There  is  a  condition,  how- 
ever, where  there  is  no  tendency  for  such  fatal  terminations, 
and  yet  the  utility  of  the  animal  is  greatly  handicapped  for 
life,  and  the  monetary  losses  to  the  owner  are  serious. 
We  all  know  how  liable  hunter  stock  are  to  become  roarers 
and  whistlers,  particularly  if  they  should  suffer  from 
influenza  or  strangles ,  and  even  severe  colds. 


156  CLINICAL  BACTERIOLOGY  AND 

Young  horses  at  an  age  when  their  dentition  processes 
are  in  full  activity  are  congregated  at  fairs  and  sales, 
come  in  contact  with  a  focus  of  infection,  and  are  suitable 
prey  for  the  many  pathogenic  bacteria  to  locate  themselves 
on  the  respiratory  mucosae.  During  these  eruptive  dental 
periods  the  natural  protective  elements  of  the  system  are 
depleted,  partly  through  malnutrition  consequent  upon 
indigestion  due  to  improper  mastication  of  the  food,  the 
phagocytes  and  the  opsonins,  etc.,  are  below  the  normal 
healthy  standard,  and  in  the  case  of  the  Irish  hunter  sent 
over  to  England  the  nervous  and  systemic  depression 
following  upon  a  long  journey  and  sudden  climatic  changes 
— not  to  speak  of  the  ill  effects  of  bad  boat  and  rail 
travelling — make  these  animals  extremely  susceptible  to 
all  kinds  of  bacterial  invasions  in  general,  but  pulmonary 
in  particular. 

Here,  then,  the  immunizer  has  opportunities  of  rendering 
good  service  to  the  seller  and  buyer  of  hunter  and  blood 
stock. 

One  cannot  deny  that  our  present  knowledge  will 
not  permit  the  use  of  a  perfect  prophylactic  sero-vaccine 
as  long  as  the  bacteriology  of  the  diseases  themselves, 
which  are  productive  of  roaring  and  whistling,  is  in  an 
imperfect  state.  Nevertheless,  this  fact  does  not  prevent 
us  from  using  the  knowledge  we  do  possess  and  turning 
it  to  good  account. 

Careful  cultivation  of  the  bacteria  found  in  the  discharges 
of  animals  suffering  from  influenza  or  strangles,  and  also 
from  those  obtained  low  down  in  the  trachea  and  the 
smaller  bronchi,  should  be  made. 

These  also  should  be  procured  from  animals  located  in 
various  centres  in  the  kingdom.  In  this  manner  not  only 
would  several  strains  of  bacteria  be  formed,  but  varying 
degrees  of  virulence  would  be  obtained,  and  the  mixing 
of  them  together  would  give  a  polyvalent  sero-vaccine. 
This  process,  of  course,  would  be  too  elaborate  and  ex- 
pensive for  individual  effort  to  carry  out,  but  it  would  be 


VACCINE-THERAPY  157 

well    worth    considering    by    horse-breeding    societies    or 
Government  departments. 

The  individual  immunizer,  however,  can  do  much  for  his 
client,  by  using  prophylactic  sero-vaccines  when  influenza  or 
strangles  has  broken  out  in  a  stud  of  young  blood  or 
hunter  stock,  to  reduce  the  risk  of  roaring  to  a  minimum. 
Every  animal  should  have  a  dose  at  the  first  appearance  of 
the  outbreak  in  a  stud,  and  this  should  be  repeated  in  three 
to  five  days,  two  to  four  injections  being  sufficient,  according 
to  the  severity  of  the  outbreak.      (See  Appendix  II.) 

Glanders. 

Mention  of  this  disease  is  made  here,  not  with  the  idea  of 
suggesting  the  use  of  a  curative  vaccine,  but  to  note  in 
passing  the  diagnostic  value  of  mallein. 

The  toxin  is  prepared  by  growing  the  bacilli  on  glycerin 
veal  for  a  month  or  six  weeks  at  37°  C.  in  flat-bottomed 
flasks,  by  which  time  a  thick  yellow  surface  growth  has 
developed. 

The  bacteria  are  then  killed  by  steaming  at  a  temperature 
of  115°  C.  for  half  an  hour. 

The  fluid  emulsion  is  next  concentrated  to  one-fourth  its 
original  volume  by  evaporation,  and  finally  filtered  through 
porcelain  to  remove  the  dead  bacilli,  after  which  it  is 
mixed  with  an  equal  volume  of  a  I  per  cent,  solution  of 
carbolic  acid  and  bottled,  and  kept  in  a  cool,  dark  room. 

If  mallein  is  injected  into  a  glandered  horse,  a  reaction 
occurs. 

The  temperature  should  be  taken  on  the  twelfth,  fifteenth, 
eighteenth,  and  twenty-fourth  hour  after  injection,  having 
been  previously  taken  once  at  least,  or  better  twice,  before 
injection. 

Should  the  temperature  rise  2' 7°  F.,  the  animal  may 
safely  be  branded  as  suffering  from  glanders,  particularly 
if  there  is  in  addition  a  local  reaction  in  the  shape  of  a 
swelling  at  the  seat  of  injection.  This  swelling  will  show 
itself  within  twenty-four  hours  of  the  injection,  and  persists 


158  CLINICAL  BACTERIOLOGY  AND 

for  three  or  four  days.  If  the  case  is  a  certain  one,  the 
swelling  will  be  at  least  4  inches  in  diameter,  and  may  be 
double,  and  is  very  painful.  If  the  animal  is  not  glandered, 
a  small  local  swelling  may  take  place  at  the  seat  of  in- 
jection, but  this  soon  subsides. 

WidaPs  agglutination  test  for  diagnostic  purposes  has 
been  extensively  carried  out  on  the  Continent,  and  in 
those  cases  where  the  temperature  is  persistently  high  it 
would  appear  this  means  of  diagnosis  is  a  useful  adjunct. 

Canine  Distemper. 

Wherever  a  canine  population  exists,  distemper  in  some 
form  or  other  is  to  be  found,  and  as  children  are  supposed 
to  have  measles  before  they  reach  maturity,  so  also  dogs 
are  expected  to  develop  distemper  during  their  early  life. 

In  approaching  the  subject  of  the  bacteriology  of  dis- 
temper, one  is  struck  with  the  number  of  alleged  causative 
bacteria  which  have  been  isolated  by  various  workers. 

Rale  in  1883  isolated  a  staphylococcus  from  pustules, 
nasal  discharges,  conjunctival  secretions,  the  internal  organs, 
and  the  blood. 

Vallerio  in  1896  discovered  an  ovoid  bacillus  in  the 
brain,  spinal  cord,  nasal  sinus,  and  conjunctival  discharges, 
and  named  it  Bacillus  caniculse. 

Piorkowski  in  1905  detected  a  bacillus  similar  to  Vallerio. 

Copeman  in  1909  discovered  a  bacillus  of  the  cocci 
group,  which  he  grew  on  gelatin  and  potato,  and  produced 
symptoms  characteristic  of  distemper  by  injecting  1  c.c. 
of  a  week-old  broth  culture. 

Lignieres  about  this  period  associated  the  causative 
organism  with  the  pasteurella  group. 

Carre,  on  the  other  hand,  claims  that  the  causative 
bacterium  of  distemper  belongs  to  the  ultravisible  group. 

Ferry,  in  1910,  adopted  a  system  of  inquiry  contrary  to 
those  of  the  previous  investigators,  inasmuch  as  he  began 
investigating  the  nature  of  the  disease  in  its  initial  stage, 
and  isolated  from  the  small  bronchi — and  sometimes  from 


VACCINE-THERAPY 


159 


the  trachea — a  short,  narrow  bacillus,  appearing  singly, 
sometimes  in  pairs,  motile,  and  aerobic. 

On  agar  plates,  after  twenty-four  hours'  inoculation  at 
37°  C,  small  colonies  appeared  slightly  raised  and  trans- 
lucent ;  after  forty-eight  hours  they  increased  in  size,  were 
curved  and  amorphous,  and  in  a  week  the  edges  undulated, 
and  showed  a  grumose  centre. 

On  potato  it  grew  with  a  characteristic  dark  brown  colour. 

On  litmus  milk,  after  seventy-two  hours,  it  grew,  giving 
the  upper  half  a  bright  blue  colour,  and  in  five  days  the 
whole  tube  was  coloured. 

Ferry  also  found  the  serum  taken  from  distemper  dogs 
always  agglutinated  this  bacillus.  To  this  organism  he 
gave  the  name  Bacillus  bronchosepticus. 

M'Gowan,  working  independently  of  Ferry,  isolated  from 
distemper  subjects  what  appears  to  be  the  same  bacillus. 
M'Gowan  believes  the  nasal  mucosa?  is  the  primary  seat 
of  development  of  the  B.  bronchosepticus,  while  Ferry  is  of 
opinion  it  settles  on  the  tracheal  mucous  membrane  at  the 
outset.     Torry  and  Hake  also  agree  with  Ferry  on  this  point. 

These  two  latter  workers,  in  addition,  give  an  interesting 
table  of  the  relative  frequency  of  the  B.  bronchosepticus  in  the 
rgans  and  fluids  of  dogs  suffering  from  distemper. 


vnriuus  or! 


1 

1 

Per  Cent,  of 

Per  Cent. 

Positive  Cases 

Per  Cent. 

Total  of 

from  which 

from  which 

in  which 

Localities  Cultivated. 

Oases 

B.  broncho- 

B. broncho- 

Locality 
Proved 

Cultured. 

septicus 

septicus 

Isolated. 

Isolated  in 
Pure  Culture. 

Sterile. 

Blood  

740 

5-4 

5-4 

71-6 

Pericardial  fluid 

28-0 

00 

0-0 

80-0 

Trachea 

52-0 

77-0 

80-0 

2  0 

Bronchials      

77-0 

74-0 

91-0 

6-5 

Lungs... 

89  0 

700 

95  0 

6-7 

Liver  

81-0 

80-0 

84-0 

30-0 

Spleen             

77-0 

17-0 

100-0 

44-0 

Kidneys 

74-0 

160 

1000 

43  0 

Nose 

68-0 

560 

23-7 

73 

Eyes 

46-0 

2-0 

o-o 

4-3 

Brain 

5-0 

20  0 

o-o 

o-o 

160  CLINICAL  BACTERIOLOGY  AND 

From  an  analysis  of  this  table  one  sees  the  B.  broncho- 
septicus  does  not  confine  itself  to  the  respiratory  tract,  and 
it  is  interesting  to  note  the  high  percentage  of  cases  in 
which  the  liver  was  involved. 

These  investigators  have  certainly  advanced  the  bacteri- 
ology of  distemper  further  than  any  other  workers,  more 
particularly  if  the  B.  bronchosepticus  proves  to  be  the 
causative  bacterium  of  distemper  proper.  In  discussing 
equine  influenza  we  expressed  an  opinion  that  the  primary 
causative  bacterium  was  probably  of  a  more  or  less  benign 
character  so  far  as  its  pathogenesis  goes,  secondary  infection 
being  responsible  for  the  complications  and  consequent 
often  fatal  sequelae. 

Canine  distemper  resembles  equine  influenza  in  many 
ways,  and  so  far  as  mixed  secondary  infections  are  con- 
cerned, it  appears  to  stand  exactly  parallel. 

The  writer  believes  also  that  the  specific  primary  infection 
is  seldom  responsible,  as  with  influenza,  for  fatal  termina- 
tions, and  if  secondary  invasions  could  be  warded  against, 
canine  distemper  would  lose  many  of  its  serious  aspects. 

The  most  common  bacteria  found  in  the  secondary 
infection  are  streptococci,  staphylococci,  and  B.  coli  com- 
munis. 

It  appears  to  be  evident,  from  the  research  work  of  Ferry, 
that  the  B.  bronchosepticus  and  its  toxins  have  such  an 
effect  upon  the  system  of  the  patient  that  the  secondary 
invading  bacteria  have  their  way  prepared  for  them,  and 
their  development  is  a  very  rapid,  virulent,  and  assured  one. 

If  this  hypothesis  is  correct,  it  will  follow,  if  Ave  desire 
to  combat  the  ravages  of  these  various  bacteria,  that  a 
vaccine  prepared  from  the  B.  bronchos epticus  must  be 
given  in  the  early  stages  of  the  disease — in  fact,  before 
the  first  manifestations  of  sickening  present  themselves, 
and  also  before  the  least  suspicion  of  a  mixed  infection 
has  taken  place,  if  benefit  is  to  follow  its  administration; 
for  it  is  obvious  that  such  a  vaccine  would  possess  no 
curative  value  whatever  against  the  destructive  effects  of, 


VACCINE-THERAPY  161 

for  example,  a  streptococcal  infection.  It  is  quite  possible 
the  failures  recorded  against  Ferry's  vaccine  are  due  to 
the  fact  that  they  have  not  been  used  soon  enough  in 
the  course  of  the  disease. 

When  secondary  infection  has  taken  place,  the  im- 
munizer  must  ascertain  in  the  first  instance  the  exact 
nature  of  the  infection ;  and,  we  believe,  owing  to  the  vari- 
able nature  of  these  infections,  to  obtain  the  maximum 
benefit  from  sero-vaccine  therapy,  the  immunizer  must  treat 
each  outbreak  or  case  on  its  own  merits,  and  for  preference 
making  his  own  autogenous  vaccines. 

In  the  past  a  variety  of  prophylactic  and  curative  vaccines 
have  been  put  forward,  some  being  highly  spoken  of,  and 
some  condemned,  each  having  its  own  exponents  as  well 
as  its  opponents. 

In  the  hands  of  some  the  results  have  been  good,  while 
with  others  they  have  been  unsatisfactory.  Why  should 
those  diversities  of  results  and  opinions  be  ? 

Can  they  be  due  to  any  want  of  skill  on  the  part  of  the 
immunizer  ?  This  is  most  improbable,  for  one  capable  prac- 
titioner praises  the  vaccine,  while  another  equally  capable 
condemns  it.  Is  it  not  rather  due  to  the  unsuitability  of 
the  vaccine  to  the  particular  outbreak  ?  and  would  we  not 
obtain  a  more  even  percentage  of  results  if  we  used  autog- 
enous instead  of  stock  vaccines  ?  Certainly,  if  we  did,  we 
would  be  less  empirical  in  our  actions,  and  probably  more 
successful  as  immunizers. 

When  a  stock  vaccine  succeeds,  we  must  reasonably  con- 
clude it  possesses  that  specificity  of  action  capable  of 
rousing  the  attacking  forces  against  the  specific  bacterium 
for  which  it  is  intended. 

The  following  are  the  principal  sera  and  vaccines  which 
have  been  used : 

Lignieres  cultivated  many  strains  from  a  pasteurella  he 
isolated  from  distemper  cases,  and  by  a  process  of  sub- 
culture which  he  carried  on  for  twelve  months,  he  was  able 
to  procure  a   strain  possessing  a  mild  degree  of  virulence. 

]l 


162  CLINICAL  BACTERIOLOGY  AND 

These  cultures  so  derived  are  incubated  in  broth-tubes  for 
fire  consecutive  days  at  a  temperature  of  42°  C,  and  from 
this  Vaccine  No.  I.  is  derived.  Vaccine  No.  II.  is  obtained 
in  the  same  way,  except  that  incubation  goes  on  for  two 
days  only.  The  dose  advised  by  Lignieres  is  1  c.c.  of 
Vaccine  No.  I.  to  be  injected  into  the  thigh  of  a  puppy; 
ten  days  later  Vaccine  No.  II.  is  injected  into  the  opposite 
thigh. 

Lignieres  at  a  later  date  injected  his  vaccine  as  above 
described  into  horses,  beginning  with  small  doses,  and 
increasing  them  as  toleration  became  established  at  regular 
intervals,  first  under  the  skin,  and  later  into  the  vein 
direct. 

The  immune  serum  is  then  obtained  by  bleeding  the 
animal,  collecting  the  blood  in  sterile  vessels,  allowing  it 
to  clot  on  ice,  and  the  resultant  serum  placed  in  stock 
bottles,  and  a  small  percentage  of  lysol  added. 

The  dose  is  from  10  to  20  c.c.  to  each  dog. 

Phisalix,  adopting  Lignieres'  method,  made  his  mono- 
valent vaccine,  which  he  derived  also  from  a  pasteurella 
cultivated  on  glycerine  and  broth.  The  dose  he  gives  is 
2  c.c.  to  each  dog,  repeating  the  vaccine  in  ten  days. 

Piorkowski  prepares  a  polyvalent  serum  which  is  highly 
spoken  of  by  many  clinicians,  while  others  consider  it  is 
useless  as  a  prophylactic  and  curative  agent. 

As  a  prophylactic  dose  he  advises  5  to  10  c.c,  im- 
munity, it  is  stated,  being  maintained  thereby  for  six 
months,  and  for  a  curative  dose  10  to  20  c.c.  is  recom- 
mended. 

Ferry,  after  isolating  his  microbe,  the  B.  bronchos ejpticus, 
made  a  protective  vaccine  mainly  to  prove  whether  this 
bacillus  was  the  real  causative  organism  of  distemper,  and, 
in  addition,  he  carried  out  agglutination  tests. 

The  following  is  an  extract  taken  from  his  paper,  "  The 
Cause  of  Distemper  in  Dogs,"  and  which  appeared  in  the 
New  York  Journal  for  July,  1912  : 

"Agglutination    Tests  and   Protective   Inoculations. —  In 


VACCINE-THERAPY  163 

order  to  strengthen  our  position  it  was  necessary  to  carry  out 
tests  other  than  experimental  inoculations — namely,  agglu- 
tination tests  and  protective  inoculations.  The  agglutination 
tests  were  carried  out  on  the  following  serums  :  From  dogs 
suffering  with  the  disease  spontaneously  contracted,  from 
dogs  experimentally  infected,  from  dogs  immunized  to  the 
live  organisms,  and  also  those  immunized  to  dead  cultures. 
These  were  tested  repeatedly  on  the  same  strain  and  on 
different  strains.  They  all  gave  positive  agglutinations, 
with  but  one  exception ;  in  this  case  the  dog  was  an  old 
one,  and  had  probably  suffered  with  the  disease.  The 
serum  from  the  dogs  suffering  from  the  disease  spontaneously 
produced,  as  those  found  on  the  streets,  gave  an  agglutina- 
tion ranging  from  1  in  40  to  1  in  800.  The  serum  from 
the  dogs  experimentally  infected  gave  an  agglutination 
from  1  in  100  to  1  in  600;  while  those  immunized  to  live 
and  dead  cultures,  gave  a  reaction  from  1  in  800  to  1  in 
4,000.  The  serum  which  gave  the  reaction  from  1  in  4,000 
was  tested  against  several  strains  of  the  bacillus,  and  the 
results  ran  from  1  in  600  to  1  in  6,000.  Two  dogs  were 
given  one  injection  of  the  live  organisms  each,  one  intra- 
venously, and  one  subcutaneously,  and  the  agglutination 
reactions  ranged  from  1  in  100  to  1  in  600.  Following 
the  injections  of  the  last  dogs,  the  only  marked  symptom 
was  diarrhoea.  This  again  emphasizes  the  symptom  upon 
which  I  have  previously  laid  so  much  stress.  In  all  of 
these  agglutination  tests  we  used,  as  controls,  blood  of 
dogs  which  had  never  had  the  disease.  They  invariably 
gave  negative  results. 

"  The  protective  inoculations  were  also  very  convincing, 
and  proved  to  our  entire  satisfaction  that  we  were  dealing 
with  the  specific  bacillus.  I  will  quote  from  my  second 
article  in  this  connection :  {  Forty  dogs  were  used  in  all ; 
nine  were  immunized  with  live  cultures,  while  fourteen 
were  saved  as  controls.  All  of  these  dogs  were  exposed  to 
at  least  three  dogs  suffering  with  typical  symptoms  of  the 
disease,  including  the  respiratory,  abdominal,  and  nervous 


164  CLINICAL  BACTERIOLOGY  AND 

types.  Eight  of  the  controls  died,  while  all  of  the  im- 
munized dogs  remained  well.'  These  dogs  were  all  in  the 
same  room  and  exposed  to  each  other,  so  there  was  plenty 
of  opportunity  to  contract  the  disease  from  each  other  had 
they  been  susceptible. 

"  From  a  practical  standpoint  we  have  found  these  pro- 
tective inoculations  of  very  great  value.  From  the  nature 
of  the  work  carried  on  in  our  laboratory,  it  is  necessary  to 
use  from  fifteen  to  twenty  dogs  a  week  for  one  purpose  or 
another,  and  it  has  always  been  our  experience  to  lose 
nearly  all  of  the  young  ones,  not  killed  at  once,  to  distemper. 
Since  beginning  to  give  some  of  these  dogs  protective  in- 
jections as  soon  as  they  are  received  from  the  dog-pound, 
our  experience  is  just  reversed,  and  we  are  able  to  save  a 
large  number  of  them. 

"  This  is  not  only  a  great  help  as  far  as  the  experiments 
are  concerned,  but  a  great  saving  from  a  monetary  point  of 
view,  and  incidentally  corroborates  our  previous  results. 
All  of  these  dogs  are  exposed  to  infection  before  being 
brought  to  the  laboratory,  and  many  of  them  have  already 
contracted  the  disease,  so  that  the  fact  that  we  are  able  to 
save  so  many  speaks  well  for  the  value  of  the  inoculations. 
We  give  three  injections,  with  two  or  three  days'  intervals, 
beginning  with  200,000,000  bacteria,  and  increasing  the 
dose  by  200,000,000. 

"  The  dogs  have  suffered  no  ill  effect  whatever  from  these 
protective  injections." 

Ferry  advises  as  a  prophylactic  the  following  doses  of 
devitalized  B.  bronchocanis,  with  intervals  of  from  three  to 
five  days  between. 

First  200,000,000 

Second       400,000,000 

Third         600,000,000 

The  vaccine,  of  course,  should  be  given  before  symptoms 
of  the  disease  present  themselves.  Ferry  suggests,  for  the 
best  results  to  be  obtained,  the  vaccine  should  be  given  a 


VACCINE-THERAPY  165 

month  previous  to  exposure  to  infection.  This  is  not 
always  practicable ;  but,  he  adds,  "  treatment  may  be  in- 
stituted at  the  time  of  exposure.  The  disease  will  not  always 
be  prevented  at  this  stage,  but  the  severity  of  the  symptoms, 
if  the  dog  becomes  infected,  will  be  much  decreased  and  the 
duration  of  the  disease  lessened." 

For  those  cases  where  the  disease  is  established,  Ferry 
prescribes  a  polyvalent  curative  vaccine,  beginning  with 
the  following  initial  dose  : 

Bronchosepticus 100,000,000 

Staphylococci       50,000,000 

Streptococci  25,000,000 

The  doses  are  repeated  every  third  or  fifth  day,  each 
dose  increasing  by  a  hundred  million  bronchosepticus,  and 
the  other  bacteria  pro  rata  according  to  symptoms  and 
reaction  shown.  In  mild  cases  four  doses  may  be  given, 
and  in  severe  cases  six  to  seven  doses  are  advised. 

Actinomycosis. 

This  disease  is  very  widely  spread  in  some  districts, 
particularly  in  low-lying,  damp  areas,  and  is  most  commonly 
seen  in  bovines. 

It  may  show  itself  as  a  localized  neoplasm  in  the  region 
of  the  neck,  jaw,  or  throat.  The  tongue  also  is  a  common 
seat  of  infection,  or  it  may  be  diffused,  affecting  important 
organs,  and  simulating  in  many  ways  tuberculosis. 

Under  medicinal  treatment  many  cases  made  good 
recoveries.  Potassium  iodide  or  hydrargyrum  iodium 
rubrum  exerts  an  almost  specific  action  upon  the  fungus. 

There  are,  however,  cases  when  these  drugs  appear  only 
to  check  or  keep  the  disease  in  abeyance  pro  tern.,  and  as 
soon  as  they  are  left  off  the  disease  renews  its  vigour.  It 
is  in  cases  such  as  these  that  vaccine-therapy  might  do 
good,  but  on  this  point  we  possess  no  knowledge. 

A  limited  number  of  cases  have,  however,  been  treated 
in  man  with  fairly  good  results. 


166  CLINICAL  BACTERIOLOGY  AND 

Wynn  describes  the  first  case  so  treated  in  the  British 
Medical  Journal  of  March  7,  1908,  thus  : 

"  The  infection  dates  back  at  least  twelve  months,  and 
six  months  prior  to  admission  to  hospital  extension  seems 
to  have  occurred  from  the  bronchi  to  the  lung  tissue,  and 
much  sputum  with  a   feculent    odour  was    expectorated. 
Subsequent  formation  of  an  empyema  required  operation, 
and   from   the    pus   a    pure    culture    of    streptothrix    was 
isolated,  and  a  vaccine  prepared  from  a  forty-eight-hour- 
old  agar  culture.     The  dose  employed  for  each  inoculation 
represented    O001     milligramme    of    bacterial    substance. 
Attempts  were    made    to  estimate    the   index,  which  was 
approximately  0'3  on  .Tanuary  3,  and  05  on  January  7  ; 
on  January  8  the  first  inoculation  of  O'OOl   milligramme 
was  given.     Twenty-four  hours  later  the  negative   phase 
was  apparently  over,  as  the  index  had  risen  to  0*7,  and  by 
January   16  was  1*2.     In  a  few  days  the  cough   became 
less    troublesome,  and  the  sputum  and  discharge  of  pus 
diminished  in  a  remarkable  way.     The  temperature  dropped 
from  over  100°  F.  to  normal,  and  remained  normal  for  three 
days.     Four   days   after   injection    the    discharge    had    so 
diminished  that  the  drainage-tube  was  removed.     A  slight 
rise  of  temperature  resulted,  and  on  the  18th   instant   a 
second  inoculation  of  0-001  milligramme  was  given.     Three 
days  later  temperature  was  again  normal,  and  remained  so. 
Subsequent  injections  were  given  on  February  11  and  25, 
and  March  11  and  27,  each  of  0-001  milligramme.     Th 
patient  gained   1  stone  6  pounds  in  weight,  and  the  con- 
dition on  discharge  was  a  thickened  pleura,  with  a  large, 
dry  cavity  in  the  lung.     There  was  no  sputum,  and  only 
occasionally    a    dry    cough.       The    patient    has    continued 
well." 

When  local  growths  are  operated  upon,  there  is  the 
possible  chance  of  a  recurrence.  We  venture  to  think  a 
course  of  vaccine  might  do  good  in  cases  of  this  kind,  but 
of  this  we  have  no  experience. 


PLATE  XL 


ACTINOMYCOSIS    BOVIS.  x    1000. 

(Hewlett's  "  Bacteriology.") 


T<>  face  pagi    Ldtf 


VACCINE-THERAPY  167 

Bovine  Tuberculosis. 

Clinical  and  Bacteriological  Diagnosis. — Since  Koch's 
discovery  of  the  bacillus  of  tubercle,  our  knowledge  of  this 
disease  as  it  affects  man  and  the  lower  animals  has  ever 
been  accumulative,  and  although  there  is  still  much  to 
learn,  probably  no  disease  has  been  more  fully  and  seriously 
investigated  than  tuberculosis. 

The  disease  affects  nearly  every  domesticated  animal, 
some  to  a  greater  degree  than  others,  depending  partly 
upon  the  idiosyncrasies  of  the  breed  or  strain,  and  partly 
upon  the  environment  of  the  individual,  etc.  It  is  not 
intended  here  to  discuss  tuberculosis  as  it  affects  all  the 
lower  animals,  but  to  confine  our  remarks  strictly  to  the 
disease  as  it  is  seen  in  the  bovine  species. 

Tuberculosis  may  run  an  acute  course  and  terminate 
suddenly,  or  it  may,  and  usually  does,  develop  an  insidious 
character — in  fact,  the  degree  of  virulence  in  many  cases 
is  so  slight  that  the  animal  shows  no  indication  of  a  removal 
from  the  usual  normal  healthy  standard. 

Recent  investigation  in  human  pulmonary  tuberculosis 
has  revealed  the  fact  that  the  bacillus  of  tubercle  is  not 
such  a  virulent  organism  as  the  early  bacteriologists  led  us 
to  believe — in  fact,  some  authorities  go  so  far  as  to  say  that 
if  no  secondary  infection  followed  the  primary  invasion  of 
this  bacillus,  the  disease  would  lose  its  appalling  signifi- 
cance altogether.  Certainly  this  teaching  is  in  accordance 
with  our  own  views  of  such  diseases  as  influenza  and 
distemper,  where  mixed  infections  are  the  order  of 
things. 

On  several  occasions  we  have  isolated  from  the  bronchial 
discharges  of  cattle  destroyed  after  reacting  to  the  tubercu- 
losis test  streptococci  rarely  and  staphylococci  commonly, 
and  have  used  on  a  limited  number  of  cows,  which  were 
certified  to  be  suffering  from  tuberculosis  and  which  reacted 
to  tuberculin,  a  vaccine  prepared  from  the  bronchial  and 
tracheal  discharges,  combined  with  tuberculin   and   poly- 


168  CLINICAL  BACTERIOLOGY  AND 

valent  antistreptococcal  serum  (bovine),  in  the   following 
combinations  : 

Streptococci      ...  ...  ...  250,000,000 

Staphylococci  ...  ...  ...  500,000,000 

Tuberculin         ...  ...  ...  2  c.c. 

Antistreptococcal  serum  (bovine)  ...  20  c.c. 

To  be  repeated  in  three  to  five  days. 

The  patient's  temperature  invariably  falls,  the  appetite 
improves,  and  the  respiration  becomes  slower  and  deeper;  the 
coat  regains  its  lustre,  the  condition  generally  improves,  and 
the  animal  puts  on  flesh.  When  the  disease  runs  a  very 
rapid  course,  and  the  immunizer  is  called  in  too  late,  auto- 
intoxication may  be  so  far  advanced  that  no  recovery  can 
be  looked  for  or  expected. 

We  have  been  greatly  struck  with  the  rapid  manner  the 
above  vaccine  has  cleared  up  the  bronchial  discharges. 
The  cases  which  run  a  latent  course  are  clearly  those 
where  secondary  invasion  has  either  never  taken  place, 
or,  if  it  has,  Nature's  antibodies  have  succeeded  in  resisting 
the  attack  to  a  large  extent.  We  strongly  believe  that  every 
animal  has  the  power  of  resisting  the  bacillus  of  tubercle, 
some  to  a  greater  degree  than  others.  (See  Appendix  II.) 
An  illustration  of  this  is  seen  in  the  way  Nature  isolates  and 
throws  out  an  artificial  wall  round  the  foci  of  tubercle  so 
imprisoned,  killing  them  off,  the  focus  often  ending  in  a 
caseating  or  calcifying  degenerate  mass,  and  all  that 
remains  to  show  what  has  taken  place  may  only  be  a 
cicatrix.  Should  the  bacilli,  however,  extensively  invade 
the  lungs,  these  foci  interfere  with  the  circulation  and 
respiration,  and  a  general  toxasmia  with  extreme  depression 
follows,  giving  an  excellent  opportunity  for  other  bacteria 
to  gain  a  footing.  They  in  turn  multiply  rapidly,  the  patient 
in  the  end  dying  from  acute  toxaemia  largely  due  to  a 
secondary  invasion.  When  an  animal  is  very  extensively 
diseased  with  advanced  tuberculosis,  we  know  tuberculin 


VACCINE-THERAPY  169 

may  give  no  reaction.  Might  this  fact  not  be  partly  due  to 
the  probable  condition  that  all  tubercle  bacilli  have  been 
killed  off,  not  only  by  their  own  toxins,  but  by  the  toxins 
and  other  poisons  from  the  secondary  invasion ;  and  that  the 
animal  at  this  stage  is  not  only  not  suffering  from  a  tuber- 
culosis in  the  strictest  sense,  but  from  an  acute  specific 
disease  due  to  the  secondary  bacterial  invasion  ? 

On  looking  through  the  whole  symptomatology  of  tuber- 
culosis, one  must  confess  there  does  not  exist  a  single  diag- 
nostic manifestation  which  may  be  taken  as  proof  positive  of 
the  presence  of  this  disease.  There  are,  however,  several 
symptoms  invariably  present,  which,  if  weighed  up  singly 
and  interpreted  collectively  (particularly  if  a  process  of 
what  might  be  called  symptomatic  elimination  is  adopted 
also),  will  give  to  the  trained  practitioner  strong  proof  upon 
which  to  base  his  conclusions. 

The  clinician  who  relies  upon  a  given  set  of  orthodox 
symptoms,  but  fails  to  interpret  minor  details  even  if  they 
be  unorthodox,  is  more  likely  to  make  errors  of  diagnosis 
than  the  practitioner  who  embarks  upon  his  clinical 
inquiry  with  a  more  open  mind. 

For  purposes  of  diagnosis  and  descriptive  convenience, 
we  propose  to  divide  tuberculosis  according  to  the  systems 
and  organs  it  affects,  not  forgetting  in  practice,  however, 
that  more  than  one  and  even  all  the  systems  may  be 
affected  at  the  same  time  and  in  the  same  animal : 

1.  Tuberculosis  of  the  respiratory  system. 

2.  Tuberculosis  of  the  alimentary  system. 

3.  Tuberculosis  of  the  genital  system. 

4.  Tuberculosis  of  the  nervous  system. 

5.  Tuberculosis  of  the  lymphatic  system. 

6.  Tuberculosis  of  the  mammary  system. 

Tuberculosis  of  the  Respiratory  System. 

When  a  primary  invasion  of  tubercle  bacilli  takes  place, 
probably  through  inhalation,  an  irritation  of  the  bronchial 


170  CLINICAL  BACTERIOLOGY  AND 

and  tracheal  mucosas  follows;  and  the  first  indication  to 
the  ordinary  observer  of  anything  being  amiss  is  a  short, 
dry,  strong,  and  intermittent  cough.  The  animal  at  this 
stage  may  look  well,  feed  well,  and  give,  if  a  milch  cow, 
the  usual  quantity  of  milk ;  in  fact,  we  have  even  noticed 
an  increased  supply  of  milk  at  this  stage,  probably  due 
to  an  increase  of  opsonins  thrown  into  the  blood-stream. 
As  time  goes  on  the  cough  becomes  more  pronounced,  is 
more  persistent,  and  not  so  dry;  and  also  there  is  an 
expectorate,  but,  as  this  is  usually  swallowed,  it  is  difficult 
to  detect.  One  can,  if  standing  on  the  left  side  of  the 
patient,  however,  often  see  it  pass  along  the  gullet  as  a 
bolus  after  a  fit  of  coughing. 

There  may  also  be  signs  of  pain  caused  by  the  act  of 
coughing,  as  evidenced  by  rigidity  of  the  costal  muscles 
and  the  emission  perhaps  of  a  grunt.  The  animal  now 
begins  to  lose  in  condition.  The  appetite  may  be  good, 
and  yet  the  patient  does  not  seem  to  thrive  as  she  did 
formerly,  or  the  appetite  may  be  bad,  with  resultant  falling 
away  of  flesh.  The  respiratory  sounds  may,  and  usually 
do,  reveal  a  variety  of  conditions.  Vesicular  murmurs  may 
be  either  increased  or  diminished,  or  even  absent.  Prom- 
inent bronchial  blowing  sounds  may  be  noticed.  There 
may  also  be  pronounced  crepitus  over  the  whole  pulmonary 
area.  There  are  very  decided  dry  or  moist  rales,  which 
are  materially  increased  on  exercise.  In  pulmonary  cases 
of  some  standing,  digestion  becomes  impaired,  this  doubt- 
less being  due  to  the  process  of  auto-intoxication  primarily 
depressing  the  vagus  and  leading  to  general  malnutrition,  etc. 
Fermentative  changes  now  go  on  in  the  alimentary  canal, 
as  evidenced  by  eructation  of  gas  and  foul-smelling  fasces. 
If  the  mediastinal  and  prebronchial  lymphatic  glands  are 
enlarged  and  diseased — and  they  usually  are — pressure 
on  the  oesophagus  takes  place,  rumination  is  interfered 
with,  and  eructation  of  gas  from  the  rumen  is  a  prom- 
inent symptom.  When  one  sees  this  symptom  promin- 
ently in  a  milch  cow  with  an  intermittent  and  troublesome 


VACCINE-THERAPY  171 

cough,  wasting  in  flesh,  and  general  symptoms  of  pul- 
monary lesions,  grave  suspicions  of  pulmonary  tuberculosis 
should  be  aroused.  Where  the  pharyngeal  and  laryngeal 
glands  become  involved,  difficulty  in  swallowing,  stertorous 
respiration,  protrusion  of  the  nose,  and  fulness  in  these 
regions  is  noticed.  These  may  burst  at  intervals  and  lead 
to  intermittent  discharges  from  the  nose  and  the  tem- 
porary remission  of  the  symptoms. 

Percussion  may  reveal  little  except  patchy  areas  of  dul- 
ness.  If  pleurisy  is  present,  palpation  between  the  ribs 
makes  the  animal  grunt  or  groan. 

When  the  pleura  is  acutely  involved,  as  it  invariably 
is  when  extensive  pulmonary  disease  exists — the  visceral 
pleura  through  continuity,  and  the  parietal  through  con- 
tiguity— friction  sounds  may  be  detected,  particularly  in 
the  early  stages,  although  not  always. 

As  time  goes  on  the  sounds  disappear,  and  where  large 
growths  exist,  dulness  over  these  areas  is  present.  The 
coat  in  these  latter  stages  is  dry  and  staring,  the  skin  being 
covered  with  yellow,  scaly  dandruff;  the  patient  does  not  lick 
herself ;  the  appetite  remains  in  abeyance ;  rumination  and 
lactation  are  both  suppressed;  and  persistent  fermentative 
diarrhoea  is  greatly  in  evidence.  The  eye  sinks,  and  a 
yellow  tinged  discharge  is  often  seen  from  the  inner  canthus. 
The  muzzle  is  dry  and  cold,  and  the  discharges  on  it  are  not 
licked  off  as  in  a  healthy  animal.  The  respiration  becomes 
shallow,  the  cough  weakens,  the  extremities  are  cold,  and 
the  animal,  in  a  state  of  persistent  and  progressive 
emaciation,  collapses,  is  unable  to  rise,  death  taking  place 
from  exhaustion,  consequent  upon  a  general  toxaemia  and 
inanition. 

Tuberculosis  of  the  Alimentary  System. 

Extensive  peritoneal  lesions  may  exist,  and  yet  the  animal 
shows  no  sign  of  derangement,  the  condition  being  often 
secondary  to  infection  from  some  other  centre.  Pressure 
in  an  inward  and  forward  direction  with  the  right  hand 


172  CLINICAL  BACTERIOLOGY  AND 

under  the  false  rib  will  sometimes  assist  the  clinician  in 
detecting  nodules.  Examination  per  rectum  should  be 
made  for  visceral  or  parietal  peritoneal  proliferations. 

Enlargement  of  the  abdominal  glands  should  also  be 
sought  for  per  rectum.  The  subsacral,  lumbar,  and  mesen- 
teric glands  can  all  be  felt  if  diseased  and  enlarged.  To 
the  feel  they  are  hard  nodules,  which  do  not  fluctuate  on 
pressure. 

The  liver  is  often  involved,  and  frequently  attains  to  a 
great  size.  It  can  be  felt  when  greatly  enlarged  over  the 
hepatic  region,  with  dulness  on  percussion.  The  intestines 
may  be  extensively  diseased,  and  yet  the  only  pronounced 
symptoms  may  be  persistent  diarrhcea,  alternated  with 
constipation.  The  faeces  are  usually  thin  and  watery,  con- 
taining gas  bubbles  suggestive  of  fermentation.  Sometimes 
pus  and  even  blood  or  mucus  may  be  present  in  the  fasces. 

Tuberculosis  of  the  Generative  System. 

This  is  not  uncommon  in  cows,  especially  in  those  cases 
of  generalized  tuberculosis,  and  is  characterized  by  a 
catarrhal  discharge  from  the  vulva,  which  usually  emits  a 
disagreeable  odour.  The  patient  either  does  not  conceive, 
or,  if  she  does,  abortion  takes  place  before  full  term  is 
reached.  Examination  per  rectum  will  reveal  a  distended 
womb,  which  may  be  hard  and  fibrous.  The  adjacent 
lymph  glands  are  enlarged,  sometimes  enormously,  and  are 
hard  and  lumpy  to  the  touch,  somewhat  like  bunches  of 
grapes. 

Tuberculosis  of  the  Nervous  System. 

The  diagnosis  of  tuberculosis  of  the  brain  or  its  meninges 
is  by  no  means  easy,  but  when  one  is  treating  or  examining 
a  patient  whose  symptoms  are  suggestive  of  tuberculosis — 
especially  generalized  tuberculosis  —  and  cerebral  dis- 
turbances manifest  themselves,  one  may  safely  suspect 
tubercular  infection  of  that  organ.  The  symptoms  most 
commonly  noticed  are  as  many  as  they  are  varied.      The 


VACCINE-THERAPY  1 73 

animal,  in  an  advanced  case,  may  have  a  staggering  gait. 
This  must  not  be  confused  with  the  rocking  motion  due  to 
weakness  and  inertia ;  epileptiform  movements  may  also 
be  noted — the  animal  may  shake  its  head,  or  hold  it  to  one 
side.  Blindness  may  be  total  or  partial;  one  ear  may  be 
drooped.  The  animal  may  be  excited  and  nervous,  or  the 
opposite,  much  depressed,  and  semicomatose;  muscular 
tremors  may  also  be  present,  and  there  may  be  partial  or 
complete  paralysis. 

Tuberculosis  of  the  Lymphatic  Glandular  System. 

When  the  disease  exists  in  any  organ  or  organs,  the 
neighbouring  lymphatic  glands  are  always  more  or  less 
involved.  These  glands  sometimes  swell  to  a  great  size. 
Where  the  prebronchial  and  mediastinal  lymphatic  glands 
are  infected  and  enlarged,  as  we  have  seen,  pressure  upon 
the  gullet  interferes  with  deglutition,  rumination,  and  con- 
sequently digestion.  The  mesenteric  glands  also  being 
diseased  must  interfere  with  assimilation,  and  can  some- 
times be  diagnosed  through  manipulation  per  rectum. 
When  the  laryngopharyngeal  glands  are  involved,  dis- 
tension of  that  region  is  noted;  also  difficulty  in  swallowing, 
and,  may  be,  stertorous  breathing,  with  a  more  or  less 
virulent  nasal  discharge,  protrusion  of  the  nose,  etc.  These 
glands  we  have  often  found,  by  passing  into  their  centres 
a  small,  sharp  lancet,  to  undergo  rapid  calcareous  degenera- 
tion, which,  if  the  lancet  is  rotated,  demonstrates  a  distinct 
gritty  feel,  giving  good  diagnostic  proof  of  the  nature 
of  the  condition. 

The  prescapular  gland  may  become  an  enormous  size, 
and  show  itself  as  a  floating  tumour.  When  this  growth 
has  also  been  cut  into  prior  to  or  after  removal,  the  same 
calcareous  changes  often  present  themselves. 

The  precrural  and  inguinal  glands  become  often  in- 
volved, and  in  making  a  clinical  examination  should 
be  looked  for  and  manipulated — in  fact,  enlargement 
of  any  of  the  lymphatic  glands  affords  important  assistance 


174  CLINICAL  BACTERIOLOGY  AND 

to  the  practitioner  who  may  be  called  upon  to   diagnose 
a  case  of  tuberculosis. 

Tuberculosis  of  the  Udder. 

This  condition  may  appear  as  a  single  nodule,  located  in 
one  quarter,  giving,  when  manipulated,  a  feeling  that  the 
surface  is  irregular,  or  it  may  be  composed  of  a  collection  of 
nodules,  giving  the  mass  considerable  dimensions  and  surface 
irregularity.  These  growths  are  hard,  firm,  and  not  painful 
when  pressed  upon. 

When  the  growth  is  confined  to  one  quarter,  the  adjacent 
healthy  parenchyma  of  the  gland  often  atrophies,  the  nodule 
thereby  assuming  a  more  or  less  pendulous  condition. 

Where  the  udder  is  extensively  involved,  the  lymphatic 
glands  on  the  supero-posterior  aspect  are  enlarged  so  much 
that  they  can  often  be  seen  and  invariably  felt.  The  pro- 
gress of  the  disease  is  usually  slow,  and  in  some  cases  a 
kind  of  parenchymatous  mammitis  develops,  due  perhaps 
to  some  form  of  secondary  infection  taking  place. 

The  udder  may  show  the  nodules  for  months,  and  yet  to 
all  appearances  the  milk  may  be  quite  normal,  and  upon 
careful  examination  no  bacilli  may  be  detected.  They 
may,  however,  be  present,  and  failure  to  detect  them  may 
be  due  to  the  fact  that  they  are  eliminated  in  limited 
numbers.  On  the  other  hand,  the  organ  may  appear  to  be 
quite  healthy,  and  yet  tubercle  bacilli  will  be  found  in  the 
milk,  evidently  coming  from  some  infective  centre  in  the 
system.  Also  it  may  be  well  to  note  that  the  udder  in  these 
cases  perhaps  harbours  the  disease  in  an  invisible  form,  there- 
by accounting  for  the  milk  contamination.  At  some  period, 
however,  an  alteration  in  the  quality  of  the  milk  will  take 
place.  As  time  goes  on  it  will  become  thin  and  watery, 
and  blue  in  colour ;  later  it  will  curdle,  and  then  a  mixture 
of  clot  and  dirty  light  red  water  is  all  that  can  be  milked 
off,  and  finally  nothing  more  than  a  dirty  grey  or  greyish- 
red  watery  fluid  appears. 


VACCINE-THERAPY  175 

Methods  of  collecting  the  Materia  Morbi  with  a  View- 
to  identify  the  Bacilli  of  Tuberculosis. 

When  one  is  called  upon  to  make  a  bacteriological  examina- 
tion of  a  case  of  tuberculosis,  one  must  collect  morbid 
material  reasonably  suspected  of  harbouring  the  bacilli.  In 
the  case  of  pulmonary  tuberculosis  the  sputum  is  chosen. 

In  those  animals  which  do  not  expectorate,  consider- 
able difficulty  is  experienced  in  collecting  the  sputum, 
and  various  methods  have  been  adopted.  Tracheotomy 
has  been  recommended  low  down  in  the  windpipe,  and  a 
swab  of  cotton-wool  inserted  to  remove  the  bronchial 
secretion,  or  a  trocar  may  be  inserted  between  the  tracheal 
rings,  and  a  feather  passed  through  the  cannula ;  the  animal 
coughs,  and  the  sputum  is  lodged  on  it  and  withdrawn. 
Others  recommend  covering  the  nose  and  mouth  with  a 
clean  sheet ;  the  animal  is  then  made  to  cough,  and  the 
expectorate  will  be  found  lodged  on  the  cloth.  The  writer 
finds  withdrawal  of  the  tongue,  previously  covered  with  a 
handkerchief  to  prevent  it  from  slipping  from  the  hand, 
and  pressing  sharply  the  larynx  to  make  the  animal  cough, 
expels  the  discharge,  which  can  be  collected  into  a  small 
plate,  easily  sterilized  by  previous  boiling.  In  some  cases 
all  we  find  necessary  is  to  withdraw  the  tongue  as  far  out 
of  the  mouth  as  possible  and  make  the  animal  cough.  The 
expectorate  will  then  usually  lodge  on  the  roof  of  the 
mouth  or  soft  palate.  The  flow  of  saliva  will  in  a  minute 
or  so  dislodge  the  expectorate,  and  if  the  nose  is  now 
depressed  the  saliva  will  float  it  on,  and  in  dropping  from 
the  mouth  it  can  be  caught  in  a  sterile  vessel. 

When  a  gland  or  glands  are  affected,  they  may  either  be 
dissected  en  masse,  or  a  piece  may  be  removed  for  micro- 
scopical examination  of  the  scrapings  therefrom.  When 
suppuration  is  in  evidence,  aspirating  the  fluid  may  answer 
the  purpose. 

In  the  case  of  mammary  tuberculosis,  the  late  Professor 
Walley  years  ago  recommended  the  use  of  a  harpoon, 
which  is  inserted  into  the  suspicious  nodule,  and  a  part  of 


176  CLINICAL  BACTERIOLOGY  AND 

the  tissue  removed  with  due  aseptic  precautions.  When  the 
milk  is  suspected  of  containing  tubercle  bacilli,  some  of  it 
should  be  stripped  from  the  quarter  and  discarded.  Brittle- 
bank  then  advises  pressing  the  udder  thoroughly,  and 
collecting  the  milk  in  a  sterile  vessel.  This  should  be 
centrifugalized  for  fifteen  minutes,  the  supernatant  fluid 
removed,  and  the  sediment  examined  microscopical^.* 

Within  recent  years  antiformin  and  other  substances 
have  assisted  investigators  in  the  isolation  of  the  bacilli  in 
pathological  fluids.  It  is  a  disinfectant  mixture  of  the 
following  composition  : 

Solution  I.  Sodium  carbonate 
Chlorinated  lime 
Distilled  water 


Solution  II.  Sodium  hydroxide 
Distilled  water 


12  grammes. 
8       „ 
80       „ 
15  grammes. 

85       „ 


Equal  quantities  of  the  two  solutions  are  mixed  together. 

The  morbid  material  containing  the  bacteria  is  placed  in 
a  centrifugal  tube,  and  antiformin  to  about  20  per  cent,  of 
its  bulk  is  added.  The  whole  solution  is  then  well  shaken, 
plugged,  and  put  in  the  dark  for  twenty-four  hours,  after 
which  it  is  centrifugalized  and  the  supernatant  fluid 
pipetted  off.  Normal  saline  solution  is  then  added  and 
the  whole  again  centrifugalized,  the  top  fluid  pipetted  off, 
and  a  loopful  of  the  remaining  sediment  is  taken  up  for 
the  purpose  of  inoculating  a  serum  tube  slope. 

The  antiformin  is  credited  with  the  power  of  destroying 
all  non-acid-fast  bacteria,  dissolving  out  mucus,  corpuscles, 
fat  cells,  etc.,  while  the  tubercle  bacilli  are  left  with  all 
their  vitality  unaffected.  The  examination  of  the  faeces  or 
the  urine  for  the  detection  of  the  bacilli  is  by  no  means 
satisfactory,  and  should  not  be  relied  upon. 

*  The  author  has  devised  an  instrument  the  object  of  which  is  to 
aspirate  the  milk  from  the  gland  through  the  teat.  In  this  way  many 
bacilli  lodged  on  or  in  the  mucosae  are  sucked  up,  thereby  increasing 
their  number  in  a  given  sample  of  milk. 


PLATE  XII. 


TUBEUCLE    BACILLI    IX    SPUTUM.        X   1000. 
(Hewlett's  "  Bacteriology.'  ) 


To  face  pagi    I  76, 


VACCINE-THERAPY  177 

The  Bacillus  of  Tuberculosis. 

This  bacillus,  in  conjunction  with  B.  lepra?,  Johne's 
bacillus,  and  certain  non-pathogenic  bacteria  isolated  from 
freces,  hay,  milk,  butter,  and  other  sources,  have  common 
staining  and  morphological  characters,  which  associate 
them  under  one  group,  known  as  the  "acid-fast  group." 
As  the  term  implies,  they  possess  the  power  of  retaining 
stains  which  resist  the  decolorizing  action  of  strong  acids. 
They  do  not  stain  readily  with  ordinary  aniline  dyes,  but 
when  stained,  they  become  "  acid- fast," 

The  members  of  this  group  are  all  slender  rods,  non- 
motile,  having  neither  capsules  nor  spores,  and  are  Gram- 
positive. 

Their  slight  variations,  morphologically  and  culturally, 
and  their  considerable  differences,  so  far  as  their  patho- 
genesis goes,  serve  to  differentiate  the  various  species. 

The  B.  tuberculosis  is  a  slender  rod,  frequently  bent,  with 
rounded  ends.  In  length  it  varies  from  0*2  to  3"5  yu,.  It  may 
stain  irregularly,  giving  the  bacillus  a  beaded  appearance. 

Save  for  Johne's  bacillus,  the  other  bacteria  of  the  acid- 
fast  group  are  non-pathogenic  to  animals,  and  their  presence 
in  animal  tissues  may  be  defined  as  accidental.  It  is  probable 
they  are  more  commonly  present  as  such  than  we  are  inclined 
to  believe.  In  any  case,  this  probable  fact  should  make  us 
the  more  careful  in  drawing  our  conclusions  and  giving 
our  opinions.  Where  one  is  permitted  to  do  so  by  law,  and 
time  for  giving  an  opinion  is  unlimited,  the  presence  of 
tubercle  bacilli  in  morbid  material  can  be  most  satisfactorily 
settled  by  the  biological  test. 

Methods  for  staining  the  Bacillus  of  Tubercle. 

Ziehl-Neelsen' s  Method. — The  materia  morbi  having  been 
collected  with  the  usual  careful  precautions  to  prevent 
outside  bacterial  contamination,  and  subjected  to  the 
methods  necessary  to  facilitate  the  easy  finding  of  the 
bacteria,  as  already  described,  a  loopful  of  the  material  is 
taken  up   by  a  sterile  platinum  loop,  and  spread  by  the 

12 


178  CLINICAL  BACTERIOLOGY  AND 

needle  over  a  grease-free  slide.  If  the  film  is  too  thin,  it 
should  be  allowed  to  air-dry,  and  another  layer  spread  on 
the  top  of  it,  this  process  being  repeated  if  necessary. 
In  the  examination  of  urine  or  of  the  watery  secretion  of 
a  badly  diseased  udder,  this  is  sometimes  needful. 

When  the  film  has  become  dry,  it  should  be  passed  two  or 
three  times  through  the  flame  to  fix  it. 

The  slide  is  then  placed  upon  a  metal  tray  under- 
neath which  is  a  lighted  spirit  lamp,  and  carbol-fuchsin 
filtered  on  to  the  specimen. 

As  soon  as  steam  begins  to  rise  from  the  slide,  it  should 
be  removed  and  decolorized  by  dipping  in  a  25  per  cent, 
solution  of  sulphuric  acid  for  three  or  four  minutes.  This 
has  the  effect  of  removing  the  stain  from  everything  except 
the  acid-fast  bacilli.  The  whole  film  now  shows  a  yellow 
tinge.  It  is  then  washed  in  water,  when  a  pale  red  colour 
returns  to  it. 

Wash  in  alcohol  until  no  more  stain  comes  away.  By 
doing  so  the  stain  is  removed  sometimes  from  acid  -  fast 
bacilli  other  than  tubercle,  if  present,  which  is  of  course  to 
be  desired. 

Wash  in  water,  counter-stain  in  weak  methylene  blue 
for  one  minute.  Wash  again  in  water,  and  then  dry 
and  mount. 

By  this  method  the  acid-fast  bacilli  are  stained  red, 
while  the  non-acid-fast  bacteria,  tissue  cells,  leucocytes,  etc., 
are  stained  blue  after  the  contrast  stain. 

Pappenheim's  Method.  —  This  method  is  credited  with 
the  power  of  differentiating  between  the  bacillus  of  tuber- 
culosis and  other  acid-fast  bacteria. 

The  specimen  is  prepared  in  the  usual  way,  and  stained 
with  filtered  carbol-fuchsin  without  heat  for  three  minutes. 
The  stain  is  now  poured  off  the  specimen,  and  corallin 
solution  applied  ;  excess  of  the  fluid  is  then  poured  off  and 
another  application  made.  Three  or  four  applications  in 
all  should  be  used.  It  is  then  washed  in  water,  dried,  and 
mounted. 


VACCINE-THERAPY  179 

The  solution  is  made  thus  : 

Corallin  ...  ...  ...  ...      1  gramme. 

Methylene   blue  (saturated  alcoholic 

solution)       ...  ...  ...  ...     120  c.c. 

Glycerin  ...  ...  ...  ...     20  c.c. 

Tuberculin. 

In  1890  Koch  introduced  his  presumed  cure  for  human 
consumption — namely,  tuberculin,  which  is  prepared  as 
follows :  A  flat-bottomed  flask  containing  4  per  cent, 
glycerin-broth  is  inoculated  with  active  B.  tuberculosis 
over  the  surface,  and  incubated  at  37°  C.  for  four  weeks. 
At  the  end  of  this  period  a  heavy  wrinkled  growth 
appears,  and  in  two  months  the  growth  is  ready  for  the 
preparation  of  the  tuberculin. 

The  contents  of  several  flasks  prepared  as  above  are 
placed  in  a  porcelain  evaporating  dish  on  a  water-bath,  and 
concentrated  to  about  a  tenth  of  its  original  bulk,  when 
the  glycerin  content  becomes  about  40  per  cent. 

Several  varieties  of  tuberculin  are  prepared  by  other 
workers,  based  upon  Koch's  original  principle,  and  which 
are  used  in  bovine  practice,  not  as  a  curative,  but  as  a 
diagnostic  agent. 

Tuberculin  as  used  for  a  Test  for  Tuberculosis. 

Tuberculin  has  proved  of  great  value  in  assisting  the 
clinician  to  diagnose  tuberculosis  in  the  lower  animals. 

The  following  are  the  principal  methods  in  use  : 

The  subcutaneous  test. 

The  ophthalmic  test. 

The  cutaneous  test. 

The  Subcutaneous  Test. — The  subcutaneous  injection  of 
tuberculin  is  by  far  the  most  satisfactory  means  of  diagnosis 
in  veterinary  practice. 

The  cervical  or  scapular  region  is  the  usual  site  for 
injection.     The  hair  should  be  clipped  over  the  part,  and 


180  CLINICAL  BACTERIOLOGY  AND 

tincture  of  iodine  well  rubbed  into  the  skin ;  and  a  dose 
of  from  2  to  3  c.c,  according  to  size  and  age  of  animal,  of 
tuberculin  injected.  The  temperature  should  be  taken 
at  least  once  before  injection,  and  on  the  ninth,  twelfth, 
fifteenth,  and  eighteenth  hours  after  injection. 

The  animal  should  be  comfortably  housed,  moderately 
fed,  and  in  cold  weather  allowed  chilled  water  only  to 
drink.  Animals  in  season,  in  advanced  pregnancy,  or  when 
suffering  from  other  diseases,  or  with  high  temperatures, 
should  not  be  tested  until  these  conditions  disappear.  It 
must  be  remembered,  in  estimating  reactions,  that  young 
animals  have  a  slightly  higher  normal  temperature  than 
adults.  When  the  temperature  rises  2- 7°  F.  above  the 
average  pre-injection  temperature,  particularly  if  the  rise 
should  go  beyond  103*3°  F.  on  the  index,  one  may  safely 
conclude  the  animal  is  tuberculous. 

In  very  advanced  cases  of  generalized  tuberculosis,  one 
may  obtain  no  reaction  whatever  to  the  test;  but  the 
appearance  clinically  of  such  cases  should  arouse  grave 
suspicions  in  the  mind  of  the  clinician,  which,  with  a  know- 
ledge of  the  above  fact,  will  prevent  him  falling  into  the 
"  no  reaction,  no  tuberculosis  "  fallacy.  On  the  other  hand, 
an  animal  may  give  a  diagnostic  reaction,  and  on  post- 
mortem reveal  no  macroscopical  indication  of  tubercular 
lesions.  Such  cases  require  very  careful  and  systematic 
investigation,  for  the  infective  centre  may  be  so  very  small 
as  to  escape  notice.  The  various  organs  may  be  apparently 
healthy,  and  a  small  gland  or  chain  of  glands  harbour  the 
disease  in  its  mildest  form.  It  is  cases  such  as  these  that 
bring  the  test  into  bad  repute,  when,  owing  to  a  superficial 
examination,  an  infective  centre,  due  to  its  smallness,  is 
perhaps  overlooked. 

When  an  animal  shows  a  persistent  high  temperature, 
one  must  wait  for  it  to  fall  before  the  tuberculin  may  be 
used.  An  animal  which  has  been  injected  with  tuberculin, 
and  reacts,  ought  not  to  be  injected  again  for  at  least  a 
month ;  for  until  the  full  effects  of  the  first  injection  have 


VACCINE-THERAPY  181 

fully  passed  off  no  reaction  is  likely  to  follow  a  second  injec- 
tion. This  fact  is  sometimes  made  use  of  by  unscrupulous 
sellers,  who  inject  their  own  animals  previous  to  expert 
examination  and  injection.  This  in  itself  is  a  strong  plea 
for  the  limited  sale  of  tuberculin,  and  its  use  restricted 
to  the  hands  of  recognized  experts.* 

The  Ophthalmic  Test. — Calmette  noticed  that  a  few 
drops  of  tuberculin  applied  to  the  conjunctiva  of  a  man 
suffering  from  tuberculosis  was  productive  of  conjuncti- 
vitis, and  Vallee  showed  that  the  same  condition  took  place 
in  animals. 

A  few  drops  of  tuberculin  are  placed  in  the  conjunc- 
tival sac  by  means  of  a  sterile  glass  pipette — that 
used  for  opsonic  work  will  answer  the  purpose — and  the 
eyelids  closed  and  gently  moved  about.  When  a  reaction 
takes  place,  the  membrana  nictitans  becomes  very  red  and 
its  border  thickens ;  the  other  eye  in  itself  acts  as  a  useful 
standard  by  which  to  estimate  the  degree  of  reaction  taking 
place  in  the  tested  eye.  The  reaction  sets  in  earlier  than  by 
the  subcutaneous  method,  from  six  to  nine  hours,  and  from 
twelve  to  twenty-four  hours  a  purulent  secretion  forms  on 
the  inner  canthus,  which  appears  as  a  long,  purulent  mass 
drying  into  a  crust,  and  finally  falling  off. 

Where  one  gets  a  slight  conjunctivitis  followed  by  a 
watery  or  even  a  mucous  flow,  these  are  of  no  diagnostic 
value;  the  discharge  must  be  purulent.  A  prior  injec- 
tion of  tuberculin  has  no  effect  upon  the  reaction  in  the 
ophthalmic  test.  This  should  prove  useful  in  those  cases 
of  unscrupulous  tampering,  and  also  in  those  animals  where 
the  temperature  is  too  high  to  use  the  subcutaneous  test. 

The  Cidaneous  Test. — Pirquet  showed  that  on  inoculating 
a  small  quantity  of  tuberculin  into  the  upper  cuticle  layers 
of  a  tubercular  person,  a  local  reddening  and  swelling  took 
place,  and  Yallee  demonstrated  that  a  similar  process  took 

*  If,  however,  from  force  of  circumstances  such  a  long  time  cannot 
be  allowed,  a  reaction  may  be  obtained  if  3  to  5  c.c.  or  more  of 
tuberculin  is  given. 


182  CLINICAL  BACTERIOLOGY  AND 

place  in  animals.  The  modus  operandi  advised  by  Vallee 
is  as  follows  : 

The  hair  is  clipped  and  the  skin  shaved  over  the  shoulder- 
blade  about  the  size  of  a  five-shilling  piece,  and  the  cutis 
scarified  with  a  sterile  lancet  and  a  50  per  cent,  or  concen- 
trated solution  of  tuberculin  painted  on  with  a  brush. 

If  the  reaction  is  a  positive  one,  a  visible  infiltration  of 
the  border  of  the  wound,  and  an  oedematous  infiltration  of 
the  area  surrounding  the  seat  of  operation,  follows,  usually 
reaching  its  maximum  on  the  second  day.  In  a  negative 
case  there  is  only  a  slight  indication  of  irritation,  due  to 
the  direct  effect  of  the  lancet.  As  it  is  advisable  to  have 
a  control  area,  a  similar  surface  of  skin  should  be  scarified 
on  the  opposite  side,  and  not  treated  with  tuberculin. 

Many  theories  have  been  put  forward  with  reference  to 
the  tuberculin  reaction ;  the  processes  which  bring  it  about 
are  probably  most  satisfactorily  based  upon  what  is  known 
as  "  anaphylaxis." 

It  has  been  found  that  injections  of  many  substances 
other  than  bacteria  and  their  products  cause  reactions  on 
the  part  of  the  tissues,  with  consequent  production  of 
antibodies.  These  substances  are  called  "antigens";  an 
antigen  is  therefore  a  tissue  stimulant,  and  the  product  of 
the  stimulation  is  an  antibody. 

Tuberculin  may  be  injected  into  a  healthy  animal,  and 
produce  no  effect  whatever ;  if  injected  into  a  tubercular 
patient,  a  reaction  follows.  There  must  be  some  great 
difference  in  the  state  of  two  such  animals,  and  the  conse- 
quent reaction  in  the  diseased  animal  is  due  to  the  probable 
fact  that  it  has  become  sensitized  against  the  bacterial 
constituents. 

This  may  also  explain  why  one  injection  of  tuberculin 
prohibits  a  reaction  to  a  second  injection  up  to  a  given 
lapse  of  time,  the  body  being  in  a  state  of  anti-anaphylaxis, 
the  sensitizing  elements  having  become  exhausted  pro  tern., 
and  requiring  time  for  their  restoration. 

When  the  tuberculin  fails  to  cause  a  reaction  in  these 


VACCINE-THERAPY  183 

cases  of  advanced  tuberculosis,  it  is  possible,  among  other 
reasons,  that  the  system  may  be  in  a  state  of  immunity  to  the 
proteins  of  the  bacillus. 


Biological  Test  for  Tuberculosis. 

Where  one  possesses  the  necessary  licence,  and  when 
the  question  of  an  extended  period  is  of  no  moment  before 
an  opinion  is  required,  the  experimental  inoculation  of  the 
morbid  material  into  a  susceptible  subject  offers  the  best 
means  of  forming  an  accurate  diagnosis. 

The  guinea-pig,  by  reason  of  its  size  and  susceptibility 
to  tuberculosis,  is  the  animal  usually  chosen  for  such  an 
investigation. 

Method. — Three  guinea-pigs  are  tested  at  the  outset  with 
tuberculin  to  assure  their  freedom  from  the  disease. 

If  milk  is  the  medium  which  is  being  investigated,  a 
quantity  should  be  milked  from  the  diseased  quarter  and 
discarded,  after  which  about  a  pint  should  be  collected, 
with  great  care,  into  a  sterile  vessel,  and  poured  into  sterile 
tubes  and  centrifugalized  for  half  an  hour  at  a  speed  of 
2,500  to  3,000  revolutions  per  minute. 

At  the  end  of  this  time  a  semi-solid  layer  of  cream  will 
have  collected  on  the  top.  Under  this  is  a  layer  of  separated 
milk,  thin  and  watery,  and  at  the  bottom  a  sediment  con- 
taining cells,  debris,  and  bacteria.  Remove  the  cream  clot, 
which  looks  like  a  plug,  and  with  a  sterile  pipette  with- 
draw all  the  liquid  from  the  tube,  leaving  only  1  or  2  c.c. 
of  the  sediment.  Triturate  this  sediment  thoroughly  and 
add  10  c.c.  of  normal  saline  solution,  and  centrifugalize 
again  for  from  ten  to  fifteen  minutes.  The  supernatant 
fluid  should  now  be  pipetted  off,  and  film  specimens  made 
of  the  sediment  in  order  to  detect  the  bacilli. 

The  cream  should  also  be  examined  in  a  similar  manner, 
as  bacilli  are  sometimes  found  in  this  medium. 

The  experimental  animals  should  now  be  inoculated  with 
about  1  c.c.  of  the  sediment  or  cream,  usually  on  the  inner 


184  CLINICAL  BACTERIOLOGY  AND 

aspect  of  the  bend  of  the  knee,  with,  of  course,  due  anti- 
septic precautions. 

Each  animal  is  weighed  daily.  At  the  end  of  the 
second  week  one  is  killed,  and  a  careful  post-mortem 
examination  made.  If  no  tubercular  lesions  are  shown, 
kill  another  at  the  end  of  the  third  week,  and  if  the  results 
are  negative  the  third  should  be  kept  alive  for  six  weeks, 
and  then  killed. 

If  lesions  looking  like  tuberculosis  exist,  a  careful  micro- 
scopical examination  for  the  acid-fast  bacilli  should  be 
made,  and  this,  combined  with  the  macroscopical  appear- 
ances, forms  conclusive  proof. 

Macroscopical  Post-Mortem  Lesions  in  Bovines. 

Having  made  a  careful  clinical  examination  of  our  sub- 
ject, and  suspecting  tuberculosis,  we  subject  her  to  the 
test  (tuberculin),  and  if  she  reacts,  and  more  particularly  if 
we  have  isolated  from  the  materia  morbi  an  acid-fast  bacillus, 
we  may  with  safety  certify  her  to  be  suffering  from  tuber- 
culosis, and  need  not  be  apprehensive — if  our  examination 
technique  is  accurate — that  our  post-mortem  investigation 
will  not  substantiate  our  ante-mortem  opinion. 

As  the  term  "  tuberculosis  "  implies,  this  disease  is  charac- 
terized by  the  formation  of  tubercles  or  nodules,  which 
tend  to  undergo  cheesy  degenerative  changes. 

In  the  early  stages  these  nodules  are  small,  grey,  clear 
growths,  but  as  they  become  older  they  take  on  a  yellow 
appearance,  like  a  piece  of  cheddar  cheese  in  colour  and 
consistence — caseation.  Later  these  nodules  often  become 
gritty  in  consistence — calcification. 

When  the  respiratory  system  is  affected,  the  disease  shows 
itself  either  as  a  diffused  deposition  of  a  large  collection  of 
small  foci,  on  section,  in  the  lung  parenchyma — miliary 
tuberculosis — or  large  centres  of  purulent,  cheesy,  or  cal- 
careous masses  may  be  distributed  through  the  lung-  tissues. 
The  mediastinal  lymphatic  glands  in  such  cases  are  usually 


VACCINE-THERAPY  185 

extensively  involved.  On  the  contrary,  no  macroscopical 
lesions  may  be  present  in  the  lungs  themselves,  while  the 
glands  may  be  affected. 

Extension  often  takes  place  to  the  pleura,  and  adhesion  of 
the  parietal  and  visceral  layers  are  common.  Characteristic 
grey  nodules  are  also  common  on  the  pleural  membrane, 
appearing  as  grape-like  growths,  being  the  condition  known 
among  butchers,  etc.,  as  "  grapes."  When  the  glands  of  the 
throat  are  affected,  they  appear  as  circumscribed  growths, 
and  when  cut  into  usually  emit  a  grey  creamy  pus  mixed 
with  calcareous  deposits  which  grate  against  the  knife. 

The  abdominal  organs  are  also  often  extensively  diseased. 

The  intestinal  mucosa,  particularly  in  young  calves 
reared  upon  tubercular  milk,  shows  ulceration,  and  in 
addition,  particularly  in  adult  animals,  the  mesenteric 
glands  become  diseased,  are  enlarged,  caseous,  and,  later, 
calcareous. 

The  liver  is  commonly  the  seat  of  tubercular  nodules,  and 
sometimes  attains  to  an  enormous  size.  These  nodules  are 
generally  full  of  pus. 

The  spleen,  also,  and  kidneys  may  be  similarly  affected. 

The  generative  organs  also  become  involved.  In  the  cow 
the  uterus  and  Fallopian  tubes  may  contain  a  huge  mass  of 
tubercular  nodules,  giving  it  the  appearance  of  a  gravid 
uterus,  and  the  ovaries  also  may  be  extensively  diseased. 

The  mammary  gland  is  a  common  seat  of  the  disease, 
where  it  is  characterized  by  the  formation  of  one  or  more 
nodular  growths  affecting  one  or  more  quarters.  The 
formation  of  small  tubercles  scattered  through  the  paren- 
chyma, and  only  detected  upon  section  of  the  gland,  is 
commonly  seen  in  practice.  A  kind  of  interstitial  mammitis 
in  some  cases  is  set  up,  in  which  case  the  organ  becomes 
enlarged,  firm,  and  fibrous,  and  cuts  with  difficulty. 

The  lymphatic  glands  in  connection  with  the  udder  are 
always  involved  and  caseous  when  the  mamma  is  diseased. 


CHAPTER  XVIII 

SWINE    FEVER 

Swine  fever  is  a  specific  contagious  and  infectious  disease 
aif  ecting  pigs  of  all  ages,  and  having  a  world-wide  distribu- 
tion. It  is  primarily  caused  by  an  ultra-visible  virus,  and 
characterized  by  inflammatory  and  necrotic  lesions  due  to 
secondary  invasions. 

This  disease,  in  common  with  several  other  diseases 
where  secondary  bacterial  invasions  play  an  important  part, 
was,  up  to  the  year  1904,  thought  to  be  due  to  a  bacillus, 
whereas  in  point  of  fact  the  bacterium  then  isolated  has 
now  been  proved  to  belong  to  the  secondary  group. 

In  that  year  Dorset  and  his  co-workers,  by  injecting 
the  blood-filtrate  taken  from  a  diseased  pig  into  a  healthy 
animal  produced  the  disease,  and  settled  once  and  for 
all  the  fact  that  swine  fever  is  caused  by  an  ultra- 
microscopic  virus. 

Symptoms. — These  are  by  no  means  constant,  nor  are 
their  interpretations  always  accurate,  the  former  largely 
depending  upon  which  causative  factor  predominates. 

Where  intestinal  lesions  are  the  most  pronounced, 
abdominal  symptoms  will  predominate.  Where  the  pul- 
monary organs  are  most  severely  affected,  thoracic  symp- 
toms will  be  evident,  and  where  both  systems  are  involved, 
a  complexity  of  symptoms  will  be  seen. 

In  the  septicemic  form  the  disease  usually  runs  a  very 
rapid  and  equally  fatal  course.  In  this  condition  the 
animal  isolates  itself,  refuses  food,  and  lies  down  con- 
stantly.    The  bowels  at  first  are  constipated,  but  this  soon 

186 


VACCINE-THERAPY  187 

gives  way  to  diarrhoea.  The  fasces  often  contain  blood 
and  mucus. 

Vomition  may  be  present,  the  vomit  often  containing 
mucus,  bile,  or  even  blood. 

Acute  purulent  inflammation  of  the  conjunctiva  is  also 
commonly  seen,  whilst  the  temperature  is  usually  high. 

In  the  abdominal  form  the  animal  isolates  itself,  refuses 
its  food,  lies  about,  usually  hidden  in  a  dark  corner. 

The  bowels,  constipated  at  first,  become  replaced  with 
diarrhoea,  the  fasces  being  very  foetid.  Emaciation  rapidly 
takes  place ;  vomition  as  a  symptom  may  be  present,  and 
the  vomit  is  often  mixed  with  bile. 

The  mucosas  of  the  mouth  and  pharynx  are  inflamed,  and 
are  often  covered  with  a  tenacious  exudate  of  a  greyish- 
yellow  colour.  Invariably  there  is  difficulty  in  deglutition, 
and  sometimes  respiration.  The  temperature  is  always 
elevated  and  the  pulse  quickened. 

The  disease  may  run  an  acute  and  rapid  course,  termi- 
nating fatally  in  a  few  days.  In  other  cases  it  may  take 
on  an  insidious  form,  and  recovery  follow  in  two  or  three 
weeks.  Such  subjects,  however,  seldom  become  valuable  for 
commercial  purposes. 

In  the  pulmonary  form  the  disease  is  characterized  in 
most  cases  by  symptoms  indicative  of  acute  pneumonia. 
The  respirations  are  hurried  and  laboured,  and  sometimes 
oral,  and  in  hot  weather,  if  the  animal  is  exerted,  it  will 
roll  over  from  sheer  exhaustion  and  a  sense  of  suffocation.  . 

To  increase  the  chest  capacity  and  facilitate  respiration, 
the  animal  will  often  sit  on  its  haunches.  The  temperature 
is  usually  very  high.  There  is  always  present  a  short, 
husky  cough,  and  a  slimy  discharge  may  be  noticed  coming 
from  the  nostrils. 

Discoloration  of  patches  of  the  skin  may  be  present. 
Emaciation  may  be  very  rapid,  death  soon  taking  place 
from  exhaustion.  On  the  other  hand,  the  disease  may  take 
on  a  chronic  phase.  The  cough  usually  persists,  with 
somewhat  laboured  respiration. 


188  CLINICAL  BACTERIOLOGY  AND 

Foetid  diarrhoea  may  be  present,  and  a  chronic  arthritis 
may  set  in  as  an  additional  symptom. 

The  animal  may  eventually  sufficiently  recover  to  put  on 
flesh,  but  seldom  thrives  well. 

The  skin  may  be  the  seat  of  pronounced  lesions  located 
under  the  abdomen  and  chest,  or  on  the  thighs,  and  in  the 
region  of  the  anus.  These  may  show  various  degrees  of 
severity,  from  red  spots  to  vesicles  and  crusts. 

In  somes  the  animal  may  be  a  subject  of  paralysis. 

Bacteriology. — The  consensus  of  opinion  seems  to  favour 
the  views  of  Garbert  and  Uhlenhuth  that  the  bacteria  in 
the  secondary  infections  in  cases  of  swine  fever  are  to  a 
greater  or  lesser  degree  normal  and  non-pathogenic  inhabi- 
tants of  the  pig.  Thus  we  find  the  Bacillus  suipestifer  in 
the  intestines,  and  the  B.  suisepticus  in  the  blood  and  lungs, 
and  it  is  only  due  to  the  debilitating  effects  of  the  ultra- 
visible  virus  that  the  bacteria  of  secondary  infection  take 
upon  themselves  an  aggressive  role.  Be  this  as  it  may,  we 
know  that  most  pronounced  lesions  are  set  up  by  the 
secondary  bacteria,  which  not  only  adds  to  the  gravity  of 
the  disease,  but  prolongs  its  course. 

It  is  presumed  the  filterable  virus  produces  a  catarrh  of 
the  intestinal  follicles,  and  while  in  this  condition  the 
B.  suipestifer  penetrates  and  multiplies  with  great  rapidity, 
producing  cellular  proliferation  and  necrosis.  At  first 
a  nodule  forms,  followed  by  an  ulcer,  and  showing  a 
necrosed  centre.  Absorption  of  the  bacteria  takes  place 
into  the  lymph-stream,  and  in  passing  through  the  mesen- 
teric glands  sets  up  a  cellular  inflammation,  and,  later,  a 
dry  caseous  necrosis. 

In  like  manner  the  B.  suisepticus  locates  itself  in  the 
circulation,  producing  a  typical  hemorrhagic  septicemia ; 
or  it  may  invade  the  lung  tissue,  causing  pneumonia,  and  in 
some  cases  the  pleura  becomes  involved,  constituting  a 
typical  pleuro-pneumonia. 

The  B.  suipestifer  belongs  to  the  paratyphus  B. 
group  :    it    is    mobile  ;    in    culture    it    appears    as    long 


VACCINE-THERAPY  189 

chains  or  even  threads.  In  the  tissues  it  is  single  or 
in  pairs. 

Stains  aniline  dyes,  Gram-negative  j  it  is  anaerobic  and 
aerobic. 

In  broth  it  becomes  turbid  with  a  resultant  sediment. 

On  gelatine  bluish  transparent  colonies  form  and  do 
not  liquefy. 

On  agar  bluish  colonies  also  appear. 

On  potato  a  brownish-yellow  growth  takes  place. 

Minimum  temperature  10°  C,  optimum  37°  C,  maximum 
40°  C. 

The  B.  suisepticus  belongs  to  the  pasteurella  group, 
and  appears  in  the  form  of  short  rods,  is  non-mobile,  and 
aerobic.  It  stains  with  the  ordinary  dyes,  and  is  Gram- 
negative. 

In  peptone  broth  the  medium  becomes  cloudy. 

On  gelatine  the  colonies  appear  like  dewdrops,  at  first 
transparent,  but  later  opaque. 

On  agar  they  grow  with  a  whitish  colour,  very  tough  in 
consistency,  and  firmly  adherent  to  the  medium. 

Minimum  temperature  15°  C,  optimum  27°  C,  maximum 
35°  C. 

Macroscopical  Post-Mortem  Lesions. 

When  swine  fever  is  investigated  in  its  septicemic  form, 
haemorrhages  occur  on  the  serous  and  mucous  surfaces,  and, 
in  addition,  serous  effusions  take  place.  The  lymphatic 
glands  are  swollen  and  infiltrated,  and  also  the  parenchyma 
of  such  organs  as  the  spleen,  liver,  and  kidneys. 

In  the  alimentary  form  the  intestines  are  most  commonly 
and  acutely  involved,  and  particularly  the  caecum  and 
large  intestines.  The  mucosae  of  the  colon  shows  round 
ulcers,  yellow  or  dark  grey  in  colour,  and  particularly 
round  the  ileo-caecal  valve,  and  extension  of  similar  ulcers 
may  take  place  to  the  adjacent  small  intestines.  In  the 
less  acute  cases  small  nodules  may  take  the  place  of  ulcers. 
These  stand  out  from  the  surface,  and  constitute  the  so- 


190  CLINICAL  BACTERIOLOGY  AND 

called  "biittms"  of  swine  fever.  In  acute  conditions  a 
typical  gastro-enteritis  with  petechial  haemorrhages  may 
be  noted. 

In  some  cases  necrosis  of  the  mucosae  over  a  large  area 
of  the  large  bowels  is  pronounced,  giving  the  surface  a 
dirty  greyish-yellow  appearance. 

The  lymphatic  glands  are  extensively  involved,  in- 
filtrated, and  often  hasmorrhagic,  and,  later,  patchy  necrosis 
is  seen. 

On  the  pharyngeal  mucosae  hasmorrhagic  patches  with 
ulceration  and  necrotic  areas  are  noted,  and  the  same  con- 
ditions may  be  seen  in  the  larynx  and  the  root  of  the 
tongue. 

Where  the  pulmonary  organs  are  involved,  we  find  a 
typical  croupous  pneumonia,  and  in  advanced  cases  multiple 
necrotic  centres  are  distributed  through  the  parenchyma 
of  the  lungs.  In  addition,  a  typical  pleurisy  may  be  present, 
which  tends  to  become  plastic,  and  sometimes  pericarditis 
is  noticed. 

Sero-Therapy. 

It  is  obvious  that  so  long  as  the  bacterium  of  swine  fever 
remains  undetected  so  long  will  it  be  impossible  to  make  a 
prophylactic  or  curative  vaccine.  To  prevent  or  cure  the 
disease  we  must  therefore  fall  back  upon  serum-therapy. 

As  far  back  as  the  year  1897  Preisz  obtained  the  blood 
from  a  pig  which  had  recovered  from  a  natural  attack  of 
swine  fever.  From  this  he  made  a  serum,  injecting  10  c.c. 
each  into  thirty  pigs,  and  produced  a  degree  of  immunity. 
These  thirty  pigs  were  placed  together  with  thirty  healthy 
pigs  and  with  a  few  affected  ones.  Eighteen  of  the  in- 
oculated pigs  sickened  and  nine  died,  while  all  of  the 
thirty  control  pigs  died. 

Since  that  period  great  improvement  in  the  technique 
has  taken  place,  and  to-day  on  the  Continent  very  great 
value  is  placed  upon  the  serum  treatment  for  swine  fever 
both  as  a  prophylactic  and  as  a  cure. 


VACCINE-THERAPY  191 

The  writer  has  recently  had  the  privilege  of  visiting  the 
State  Serum  Institute  at  Rotterdam,  and  by  the  courtesy  of 
the  Director — Dr.  Poels — a  very  full  and  detailed  descrip- 
tion of  the  modus  operandi  of  swine-fever  serum  was  gone 
into,  which  is  briefly  as  follows : 

Four  young  pigs  about  eight  weeks  old  are  each  injected 
with  2-6  c.c  of  defibrinated  virulent  blood.  When  the 
disease  is  established,  they  are  killed  and  their  blood 
collected,  the  serum  of  which  is  used  to  hyper-immune  an 
adult  pig.  Into  this  pig  1  c.c.  of  virulent  serum  with 
10  c.c.  of  immune  serum  is  injected. 

14  days  later  1  c.c.  virus,  100  c.c.  immune  serum. 

14  „  „  200  c.c. 

14  „  „  „  „ 

14  „  „  „ 

14  ,,  „  ,,  ,, 

The  pig  in  ali  has  now  had  900  c.c.  of  immune  serum, 
and  is  hyper-immune.  Dr.  Poels  emphasized  the  interest- 
ing fact  that  pig  serum  can  only  produce  antibodies 
against  swine  fever,  and  that  horse  serum  is  quite 
useless.  The  drawback  to  this  is  obvious.  The  pig  being 
a  small  animal  a  limited  quantity  of  serum  can  only  be 
obtained  at  a  reasonable  price. 

The  hyper-immune  pig  (such  pigs,  by  the  way,  must  be 
possessed  of  long  tails)  is  now  bled  to  procure  the  immune 
serum.  After  thoroughly  disinfecting  the  tail  and  buttocks, 
the  animal  is  placed  in  a  kind  of  stocks  to  keep  it  in  a 
fixed  position.  The  tail  is  then  cut  at  the  end,  and  the 
stump  placed  in  a  glass  tube  with  a  capacity  of  200  c.c, 
and  700  to  1,000  c.c.  of  blood  collected ;  seven  days  later 
the  operation  is  repeated.  As  a  rule  seven  to  fifteen 
bleedings — the  latter,  if  the  tail  is  a  long  one — can  be 
obtained  from  one  pig.  The  pig  is  then  killed  by  bleeding, 
and  from  5  to  7  litres  of  serum  is  collected.     The  serum  thus 


192 


CLINICAL  BACTERIOLOGY  AND 


togethe 


collected  from  the    tail  and    throat  is   mixed 
preservative  added  of  the  following : 

Ac.  carbolic.    ...  ...  ...       5-5  c.c. 

Glycerine         ...  ...  ...     20*0  c.c. 

Aquas  dest.      ...  ...  ...     74*5  c.c. 

100  c.c.  of  the  above  is  added  to  1  litre  of  serum,  and  is 
placed  in  10,  20,  30  c.c.  bottles  and  sealed  ready  for  use. 
The  dose  for  an  adult  pig  is  from  10  to  30  c.c. 


Fig.  37. 


-Vessel  used  at  the  Serum  Institute  for  collecting 
Blood  from  the  Tail  of  an  Immune  Pig. 


Pigs  from  five  to  ten  days  old  receive  5  c.c.  The  serum 
is  injected  subcutaneously  behind  the  ear.  In  those 
cases  where  a  pig  is  being  hyper-immunized  and  large 
quantities  of  serum  is  used,  one-half  is  injected  behind  one 
ear  and  the  other  half  behind  the  other. 

When  pigs  are  injected  with  the  immune  serum,  im- 
munity is  only  conferred  for  a  period  of  three  weeks.  This 
period,  of  course,  is  of  too  short  a  duration  to  be  of  any 
practical   use.     Should,  however,  the  pig    after   injection 


VACCINE-THERAPY  193 

become  infected  with  the  virus  in  a  natural  manner,  that 
pig  is  immune  for  life. 

It  is  therefore  imperative  for  the  successful  immunization 
of  a  herd  of  pigs  that  the  virus  of  infection  must  be  always 
present,  and  without  it  no  permanent  immunity  can  be 
conferred. 

The  writer  inquired  of  Dr.  Poels  what  his  experience 
was  of  artificial  infection  in  event  of  his  failing  to  have 
the  means  of  natural  infection — i.e.,  a  diseased  pig — at 
hand,  and  his  answer  was,  it  was  not  serviceable,  but  that 
he  was  experimenting  by  feeding  pigs  with  the  natural 
virus,  and  was  hopeful  in  time  to  overcome  the  present 
difficulty  of  no  natural  infection,  no  immunity. 

The  procedure  adopted  in  Holland  is  this  :  When  a  pig 
shows  symptoms  of  swine  fever,  the  whole  herd  is  injected 
with  serum.  The  badly  affected  pigs  are  separated  from 
the  slightly  affected  ones,  and  the  experience  of  those  in 
authority  is  very  favourable  to  serum  therapy.  In  Dr. 
Poels'  words,  "The  slightly  affected  always  recover;  the 
very  badly  not  often." 

Dr.  Von  Velseim's  experience  is  that  after  the  yearly 
injection  of  2,000  pigs,  6  to  8  per  cent.  die. 

In  Holland  there  are  no  restrictions  placed  upon  swine 
fever  areas,  no  compulsory  notification,  and  no  slaughter. 


13 


APPENDIX  I 

SERO-VACCINE  THERAPY  IN  HOLLAND  AND 
OTHER  COUNTRIES 

Seko-vaccine  therapy  has  made  greater  advances  in 
Continental  countries  than  it  has  in  Great  Britain,  for  the 
obvious  reason  that  the  respective  Governments  of  these 
countries  recognize  what  a  valuable  asset  the  principle  of 
the  treatment  is,  and  freely  place  funds  at  the  disposal  of 
experts,  who  give  their  time  and  energy  to  the  advance- 
ment of  the  science.  Those  in  authority  see  the  great  gains 
agriculture  obtains  from  scientifically  applied  preventive 
and  curative  medicine. 

Thus  we  find  institutions  erected,  laboratories  fitted  up 
and  efficiently  staffed  by  excellent  men  well  paid,  whose 
duty  it  is  to  study  animal  diseases,  carry  out  research  work, 
and  adopt  scientific  measures  in  the  treatment  of  animal 
diseases. 

Such  an  institution  is  the  State  Serum  Institute  of  the 
Dutch  Government,  which  is  doing  a  very  great  work  for 
the  agriculturist  in  particular,  and  the  whole  State  in 
general. 

The  sceptic  might  say  this  is  all  very  true,  but  sero- 
vaccine  therapy  is  a  comparatively  new  method  of  treatment 
with  few  disciples,  and  it  has  not  withstood  the  test  of  time. 

In  an  exact  sense  this  is  not  so,  for  all  animals  and 
human  beings  themselves  are  by  Nature  vaccine  therapeutists 
— nay,  more,  this  fact  is  shrouded  in  antiquity,  for  it  dates 
back  to  the  period  of  the  development  of  the  first  patho- 
genic microbe.  We  know  we  are  daily  admitting  into  our 
bodies  disease  -  producing  germs  through  ingestion  and 
inhalation,  through  abrasions  of  the  skin  and  mucosas,  and 
we  are  continually  manufacturing  in  our  economies  pro- 
tective vaccines  to  fight  against  these  deadly  bacteria. 
If  it  were  not  so,  it  is  appalling  to  think  what  the  conse- 

194 


VACCINE-THERAPY  195 

quences  would  be — most  probably  complete  annihilation  of 
the  whole  human  and  animal  race. 

The  vaccine  therapeutist  is  following  out  Nature's  own 
mode  of  treatment,  which  she  has  been  adopting  for  genera- 
tions, and  it  is  only  when,  for  a  complexity  of  reasons,  she 
fails  or  appears  to  fail  that  the  science  and  art  of  vaccine 
therapy  need  to  be  called  in;  and  so  by  stimulating  the 
depressed  antibodies  and  strengthening  them  to  overcome 
the  bacterial  invasion,  she  restores  an  infected  and  diseased 
annual  to  a  healthy  and  normal  condition. 

Again,  the  sceptic  may  look  and  obtain  food  for  thought 
at  the  rapid  progress  made  by  those  Continental  institutions. 

Take,  for  example,  the  Serum  Institution  at  Rotterdam, 
and  what  do  we  find?  It  was  established  in  the  year  1904, 
and  could  only  claim  all  told  15  horses  supplying  serum. 
To-day  there  are  150  horses,  50  cattle,  and  100  pigs  used 
in  the  production  of  serum. 

During  this  period  23,000,624  animals  have  been  treated 
by  serums  and  vaccines  supplied  from  the  institution. 
Such  figures  as  these  speak  for  themselves  and  prove  con- 
clusively that  the  institution  and  the  principle  of  treatment  it 
adopts  has  become  a  valuable  asset  and  a  national  necessity. 

At  the  present  time  the  following  sera  and  vaccines  are 
in  daily  use  : 

1.  Serum  and  vaccine  against  abortion  in  mares  and  cows. 

2.  „  „  anthrax. 

3.  „  „  quarter- evil. 

4.  „  „  swine  erysipelas. 

5.  Serum  against  contagious  pneumonia  in  horses. 

6.  „  strangles. 

7.  „  tetanus. 

8.  „  foot-and-mouth  disease. 

9.  „  white  scour  in  calves. 

10.  „  septic  pleuro-pneumonia  in  calves. 

11.  „  metritis. 

12.  „  mammitis. 

13.  „  swine  fever. 

14.  „  fowl  cholera. 

The  institute  is  divided  into  twenty -four  sections,  the 
most  important  of  which  are — 

1.  The  preparation  of  serum. 

2.  The  preparation  of  vaccines. 


196  CLINICAL  BACTERIOLOGY 

3.  The  control  of  serums  and  vaccines. 

4.  The  examination  of  the  immunity. 

5.  Inquiry  into  diseases  affecting  horses,  cattle,  sheep, 
swine. 

6.  Inquiry  into  unknown  causes  of  diseases. 

7.  The  examination  of  milk. 

8.  The  examination  of  tuberculosis  of  cattle. 

9.  The  examination  of  secret  remedies. 
10.  The  chemical  section. 

The  institution  is  presided  over  by  that  distinguished 
veterinary  surgeon,  Dr.  Poels,  and  an  able  staff  numbering 
about  fifty. 


APPENDIX  II 
ADDITIONAL  NOTES 

The  Bacteriology  of  "  Whistling  and  Roaring." 

On  p.  157  we  discussed  incidentally  the  advisability  of 
using  prophylactic  sero-vaccines  in  cases  of  influenza  and 
strangles,  with  the  view  not  only  of  saving  life  and 
shortening  the  attack,  but  also  preventing  the  occurrence 
of  roaring  as  a  sequel.  It  may  not,  therefore,  be  out  of 
place  here  to  briefly  discuss  a  phase  of  the  etiology  which, 
so  far  as  we  know,  has  not  obtained  the  prominence  it 
demands  or  the  investigation  it  deserves. 

The  anatomical  arrangement  of  the  horse's  larynx  with 
its  peculiar  nerve-supply,  the  general  conformation  of  the 
neck  and  chest,  the  exerting  character  of  the  work  hunters 
and  blood  horses  are  called  upon  to  perform,  must  ever 
remain  exciting  causes  in  the  production  of  the  conditions 
known  as  "  whistling  "  and  "  roaring  "  proper.  The  actual 
cause,  we  venture  to  think,  is  attributed  to  the  specific 
action  of  bacteria  and  their  products  in  the  majority  of  cases. 

The  inflammatory  sore  throat  one  sees  in  practice  is  due 
to  bacteria.  The  nasal,  laryngeal,  and  pharyngeal  catarrhs 
are  bacterial  in  origin;  influenza,  bronchitis,  pneumonia, 
and  pleurisy  are  all  caused  by  specific  micro-organisms, 
and  we  know  roaring  is  a  common  sequel  to  any  or  all  of 
these  conditions.  Horses  also  are  liable  to  become 
grunters  following  upon  these  conditions,  and  grunting 
invariably  is  the  precursor  of  roaring. 

The  fact  that  a  horse  grunts  when  "  ribbed "  suggests 
a  general  derangement  of  the  nervous  system,  the  nature 
of  this  derangement  being  hyper-sensitiveness,  owing  to 
malnutrition  and  a  consequent  probable  toxaemic  neuritis. 

In  addition  to  these  constitutional  disturbances  we  must 
also  look  to  local  conditions.  There  we  find  the  laryngeal 
197 


198  CLINICAL  BACTERIOLOGY  AND 

mucosae  acting  as  an  excellent  medium  upon  which  a 
rich  bacterial  growth  is  taking  place.  Consequent  upon 
this  growth  the  whole  surface  is  bathed  with  endotoxins 
and  exotoxins  derived  from  the  pathogenic  bacteria. 

Absorption  of  these  takes  place  into  the  deeper 
structures,  including  the  muscular  elements  of  the  larynx 
extending  to  the  nerve  endings,  the  nerve  fibres,  and  nerve 
trunks  themselves,  and,  according  to  the  structures 
involved,  we  now  find  a  specific  mucositis,  cellulitis, 
myositis,  and  neuritis  set  up.  This  is  followed  by 
degenerative  changes ;  the  trophic  functions  of  the  re- 
current nerve  are  soon  interfered  with,  and  muscular 
atrophy  is  the  result.  These  conditions  are  aggravated  by 
the  strain  placed  upon  the  already  depressed  set  of 
muscles,  and  the  degenerative  changes  which  have  already 
taken  place  in  the  nerve  are  increased  by  the  jerking  of 
the  aorta  round  which  the  nerve  passes. 

The  muscles  on  the  right  side  of  the  larynx  are  subject 
to  the  same  degenerative  causes,  and  no  doubt  temporary 
atrophic  changes  take  place,  but  owing  to  the  different 
disposition  of  the  nerve-supply,  the  trophic  interference  is 
not  called  into  play. 

Undoubtedly  there  are  cases  where  the  mildness  of  the 
attack  has  only  been  capable  of  producing  slight  degenera- 
tive and  organic  changes  both  in  the  muscles  and  the 
nerve,  and,  given  time,  a  complete  recovery  follows.  This 
may  explain  a  very  common  fact  we  see  in  practice — 
namely,  an  animal  recently  recovered  from  a  catarrh,  etc., 
may  be  a  pronounced  whistler,  and  even  a  roarer,  and,  in 
time,  with  exercise,  good  food,  and  conditioning,  may  be- 
come quite  sound  in  his  wind,  the  obvious  conclusion  being 
the  retrogressive  changes  were  so  slight  that  Nature  was 
able  to  bring  about  a  complete  restoration  before  hopeless 
organic  chaos  took  place. 

If  these  foregoing  conclusions  are  correct,  it  is  obvious, 
if  we  succeeded  in  preventing  bacterial  invasions  to  the 
upper  air  passages,  we  should  hear  very  little  of  roaring 
and  whistling  in  horses,  but  such  a  task,  of  course,  would 
scarcely  be  practicable.  We  can,  however,  reduce  the 
severity  of  a  bacterial  attack,  and  in  many  cases  even 
prevent  its  development  altogether  by  sero- vaccine  therapy. 

It  may  be  pointed  out  heredity  plays  an  important  part 
in  the  production  of  roarers  and  whistlers,  and  with  this, 
in  a  limited  sense,  we  agree.     Conformation  is  undoubtedly 


VACCINE-THERAPY  199 

hereditary,  and  we  know  horses  with  long  necks,  narrow 
intermaxillary  spaces,  long  backs,  narrow  chests,  etc.,  are 
all  liable  to  become  roarers.  But  this  point  does  not 
interest  us  here  so  much  as  the  fact  that  some  animals 
inherit  in  a  pronounced  degree  a  defective  resisting-power 
to  bacterial  invasion. 

We  know  some  horses  are  more  predisposed  to  the  disease- 
producing  effects  of  the  virus  of  strangles  and  influenza 
than  others,  one  animal  suffering  slightly  or  not  at  all,  and 
another  severely.  The  protective  antibodies  of  the  one  are 
more  resistant  than  in  the  other,  and  we  are  of  opinion  that 
it  is  an  inherited  deficiency  of  the  antibodies  which  largely 
accounts  for  the  practical  conclusions  of  roaring  appearing 
most  often  in  certain  strains,  and  from  one  generation  to 
another. 


The  Drain  of  Antibodies  through  the  Milk-Supply  as 
a  Probable  Predisposing  Cause  of  Tuberculosis. 

On  p.  168  we  stated  our  belief  that  every  animal  has  the 
power  of  resisting  the  bacilli  of  tubercle,  some  to  a  greater 
degree  than  others.  In  such  a  complex  mechanism  as  the 
animal  body,  provided  as  it  is  with  a  variety  of  protective 
bodies,  many  of  which  are  little  understood,  it  may  be  that 
more  than  one  antibody  is  deficient,  and  it  is  possible  a 
variety  of  forces  are  at  play  to  account  for  this  deficiency. 

We  know  milch  cows  are  more  prone  to  tuberculosis 
than  any  other  class  of  our  domesticated  animals.  More- 
over, some  strains  appear  to  be  more  predisposed  to  the 
disease  than  others,  and  we  invariably  find  the  deeper  the 
milker  the  greater  the  predisposition  to  tubercle. 

The  pertinent  question  may  here  be  asked,  Why  should 
this  be  ? 

The  depletion  of  the  system  through  the  milk-supply 
undoubtedly  accounts  for  a  lowering  of  the  vital  forces, 
and  thereby  making  the  animal  a  fit  subject  for  bacterial 
invasion.  But  why  should  the  physiological  act  of  lactation, 
and  even  excessive  lactation,  predispose  the  animal  to 
tuberculosis  ?  Such  a  normal  act  should  not  disturb  the 
metabolism,  and  make  the  subject  a  suitable  medium  for 
bacterial  growth. 

It  is,  therefore,  not  enough  to  say  the  vital  forces  are 
lowered  by  milking.     There  must  also  be  a  drain  upon  the 


200  CLINICAL  BACTERIOLOGY 

bacteriotropic  and  bacteriolytic  elements  of  the  blood, 
which  find  their  outlet  by  the  milk  stream.  We  know 
opsonins  are  to  be  found  in  the  urine  and  in  the  perspira- 
tion, and  why  not  in  the  milk,  which,  in  addition  to  being 
a  secretion,  is  also  an  excretion  up  to  a  point.  And  if  this 
is  true  of  opsonins,  why  not  the  other  antibodies  of  the 
blood  also  ?  A  system  so  depleted  of  its  protective  anti- 
bodies must  therefore  be  liable  to  bacterial  attacks,  and 
more  so  if  these  antibodies  are  specific  to  the  causative 
bacteria. 

We  have  over  and  over  again  noticed  cows  suffering 
from  tuberculosis  in  a  moderately  mild  form  give  a  larger 
volume  of  milk  per  diem  than  cows  deemed  free  from  the 
disease,  but  it  would  appear  to  be  the  quantity  is  increased 
at  the  expense  of  the  quality.  These  cows  are  generally  in 
very  poor  condition,  and  even  in  advanced  stages  of 
emaciation.  Now,  if  such  cows  are  allowed  to  become 
"  dry/'  it  is  astonishing  how  the  symptoms  common  to 
tuberculosis  abate,  and  the  animal  begins  to  thrive  and 
often  do  well,  and  in  time  an  apparent  complete  cure 
follows. 

Some  practitioners  will  explain  that  this  is  due  to  a  check 
upon  the  nutritive  material  in  the  form  of  milk  leaving  the 
system  and  disturbing  the  metabolism.  This  may  be,  and 
undoubtedly  is,  true  up  to  a  point,  but  that  fact  would  not 
in  itself  sufficiently  explain  the  improvement  taking  place 
in  the  diseased  organs.  This  improvement,  we  believe,  is 
brought  about  by  a  rise  in  the  opsonic  index  owing  to  a 
check  upon  the  outflow  of  opsonin  into  the  milk  stream ; 
and,  of  course,  an  increase  of  opsonin  in  the  system  means 
a  more  powerful  offensive  and  defensive  force  attacking  the 
invading  bacteria,  and  bringing  about  their  destruction,  and 
the  restoration  of  the  patient  to  a  more  healthy  standard. 


APPENDIX   III 
WEIGHTS  AND  MEASURES 

(a)  The  initial  unit  of  the  Metric  System  is  the  metre  (m.). 

(b)  The  unit  of  mass  is  the  gramme  (g.),  which  equals 
the  weight  of  1  cubic  centimetre  of  water  at  its  maximum 
density. 

(c)  The  unit  of  the  measure  of  capacity  is  the  litre  (1.), 
which  equals  the  volume  of  1  kilogramme  of  water  at  its 
maximum  density. 


0») 

(mm.) 

(cm.) 

(1") 


1  micron  - 
1  millimetre 
1  centimetre 
1  inch 


Length. 


inch. 


5  00  0 

K  inch 


25  millimetres. 


Mass. 


(nag.)     1  milligramme  - 
(g.)        1  gramme 
(kg.)      1  kilogramme   - 


-  =    0-01543  grain. 

-  =    15-4323  grains. 

-  =   2  pounds  3£  ounces. 


(lb.)       1  pound  (Avoirdupois) 
(oz.)       1  ounce  „ 

(gr.)      1  grain 


453-592  grammes. 
28-35  grammes. 
0-0648  gramme. 


Volume  or  Capacity. 


(c.c.) 

ao 

1  cubic  centimetre 
1  litre 

-    = 

16-9  minims. 
35-196  fluid  ounces. 

(5) 
(0.) 

1  ounce    - 
1  pint 

-    = 

28-42  cubic  centimetres. 
568-34  cub.  centimetres. 

202  CLINICAL  BACTERIOLOGY  AND 

To  convert  grammes  into  grains      -         -  x  15*432. 

,,  grammes  into  ounces     -         -  x  O03527. 

„  kilogrammes  into  pounds       -  x  2*2046. 

„  cubic  centimetres  into  ounces  x  0*0352. 

„  litres  into  fluid  ounces  -  x  35*2. 

To  convert  grains  into  grammes      -         -  x  0*0648. 

„  ounces  into  grammes     -         -  x  28*35. 

„  ounces  into  cubic  centimetres  x  28*42. 

„  pints  into  litres     -         -         -  x  0*568. 


THE1LMOMETKIC  SCALES 

On  the  Fahrenheit  scale  the  freezing-point  of  water  is 
32°,  and  boiling-point  212°;  on  the  Centigrade  scale  the 
freezing-point  is  at  zero,  and  the  boiling-point  at  100°. 

The  following  formula  is  used  for  converting  Fahrenheit 
into  Centigrade  and  vice  versa  : 

F.  into  C.  =  (degree  -  32)  -*■  9  x  5. 
C.  into  F.  =  degree  -*-  5  x  9  +  32. 


Appended  are  two  complete  tables 


VACCINE-THERAPY 


203 


TABLE  FOR  THE  CONVERSION  OF  FAHRENHEIT 
INTO  CENTIGRADE  DEGREES 


Fahr. 

Cent. 

Fahr. 

Ctnt. 

Fahr. 

Cent. 

Fahr. 

Cent. 

32 

o-o 

78 

25-6 

123 

50-6 

168 

756 

33 

0-6 

79 

26-1 

124 

51-1 

169 

761 

!  34 

11 

80 

26-7 

125 

517 

170 

76-7 

35 

1-7 

81 

27-2 

126 

52-2 

171 

77-2 

36 

2"2 

82 

27-8 

127 

52-8 

172 

77-8 

37 

2-8 

83 

28-3 

128 

53-3 

173 

78-3 

38 

3-3 

84 

28-9 

129 

53-9 

174 

78-9 

39 

3-9 

85 

29-4 

130 

54-4 

175 

794 

40 

4*4 

86 

30-0 

131 

55-0 

176 

800 

41 

5-0 

87 

30-6 

132 

556 

177 

806 

42 

5-6 

88 

31-1 

133 

56-1 

178 

811 

43 

6-1 

89 

31-7 

134 

56-7 

179 

81-7 

44 

67 

90 

32-2 

135 

57-2 

180 

82-2 

45 

7-2 

91 

328 

136 

57-8 

181 

82-8 

46 

7-8 

92 

33-3 

137 

58-3 

182 

83-3 

47 

8*3 

93 

33-9 

138 

58-9 

183 

839 

48 

8-9 

94 

34-4 

139 

59-4 

184 

84-4 

49 

9-4 

95 

350 

140 

60  0 

185 

85-0 

50 

10-0 

96 

35-6 

141 

60-6 

186 

85-6 

51 

10-6 

97 

36-1 

142 

611 

187 

861 

52 

11-1 

98 

367 

143 

61-7 

188 

86-7 

53 

11-7 

99 

37-2 

144 

62-2 

189 

87-2 

54 

122 

100 

37-8 

145 

62-8 

190 

87-8 

55 

12-8 

101 

38-3 

146 

63-3 

191 

88-3 

56 

133 

102 

38-9 

147 

63-9 

192 

88-9 

57 

139 

103 

39-4 

148 

64-4 

193 

894 

58 

144 

104 

400 

149 

65-0 

194 

900 

59 

15'0 

105 

40-6 

150 

65-6 

195 

906 

60 

15-6 

106 

41-1 

151 

66-1 

196 

911 

61 

161 

107 

41-7 

152 

66-7 

197 

917 

62 

16-7 

108 

42-2 

153 

67-2 

198 

922 

63 

17-2 

109 

42-8 

154 

67-8 

199 

92-8 

64 

17-8 

110 

43-3 

155 

683 

200 

933 

65 

183 

111 

43-9 

156 

68-9 

201 

939 

66 

18-9 

112 

44-4 

157 

69-4 

202 

944 

67 

194 

113 

45  0 

158 

70-0 

203 

950 

68 

20-0 

114 

456 

159 

70  6 

204 

956 

69 

206 

115 

46-1 

160 

71-1 

205 

961 

70 

211 

116 

46-7 

161 

71-7 

206 

967 

71 

21-7 

117 

47-2 

162 

72-2 

207 

972 

72 

22-2 

118 

47-8 

163 

72-8 

208 

97-8 

73 

22-8 

119 

48-3 

164 

73-3 

209 

98-3 

74 

233 

120 

48-9 

165 

73-9 

210 

989 

75 

239 

121 

49-4 

166 

74-4 

211 

994 

76 

24-4 

122 

500 

167 

75-0 

212 

1000 

77 

25-0 

204 


CLINICAL  BACTERIOLOGY 


TABLE  FOR  THE  CONVERSION  OF  CENTIGRADE 
INTO  FAHRENHEIT  DEGREES 


Cent. 

Fahr. 

Cent. 

Fahr. 

Cent. 

Fahr. 

0 

32-0 

34 

932 

68 

154-4 

1 

33-8 

35 

950 

69 

1562 

2 

35-6 

36 

96-8 

70 

1580 

3 

374 

37 

986 

71 

1598 

4 

39-2 

38 

1004 

72 

161-6 

5 

41-0 

39 

102-2 

73 

1634 

6 

42-8 

40 

1-040 

74 

1652 

7 

44-6 

41 

1058 

75 

1670 

8 

46-4 

42 

107-6 

76 

168-8 

9 

48-2 

43 

1094 

77 

1706 

10 

50-0 

44 

111-2 

78 

172-4 

11 

51-8 

45 

1130 

79 

174-2 

12 

53-6 

46 

1148 

80 

1760 

13 

55-4 

47 

1166 

81 

177-8 

14 

57-2 

48 

1184 

82 

179-6 

15 

59-0 

49 

1202 

83 

1814 

16 

60-8 

50 

1220 

84 

183-2 

17 

62-6 

51 

123-8 

85 

185-0 

18 

64-4 

52 

1256 

86 

186-8 

19 

66-2 

53 

1274 

87 

188-6 

20 

68-0 

54 

1292 

88 

1904 

21 

69-8 

55 

1310 

89 

192-2 

22 

71-6 

56 

132-8 

90 

1940 

23 

73-4 

57 

134-6 

91 

195-8 

24 

752 

58 

136-4 

92 

1976 

25 

77-0 

59 

138-2 

93 

1994 

26 

78-8 

60 

1400 

94 

2012 

27 

80-6 

61 

141-8 

95 

203-0 

28 

82-4 

62 

1436 

96 

204-8 

29 

84-2 

63 

145-4 

97 

2066 

30 

86-0 

64 

147-2 

98 

208-4 

31 

87-8 

65 

1490 

99 

2102 

32 

89-6 

66 

1508 

100 

2120 

33 

914 

67 

152-6 

APPENDIX  IV 


(8798.) 


ORDER  OF  THE  BOARD  OF  AGRICULTURE 
AND  FISHERIES. 

(Dated  13th  February,  1913.) 


TABLE  OF  CONTENTS. 

Article. 

1.  Interpretation. 

2.  Notice  of  Disease. 

3.  Notification  of  Disease  by  Veterinary  Surgeons. 

4.  Inspection  and  Examination  of  Animals. 

5.  Slaughter  of  Diseased  Animals. 

6.  Valuation  for  Compensation. 

7.  Post-Mortem  Examination  of  Slaughtered  Animals. 

8.  Compensation. 

9.  Precautions  to  be  adopted  with  Respect  to  Milk,  etc. 

10.  Detention  and  Isolation  of  Suspected  Animals. 

11.  Suspected  Animals  in  Markets,  Fairs,  and  Sales. 

12.  Cleansing  and  Disinfection. 

13.  Reports  to  the  Board. 

14.  Extension  of  Certain  Sections  of  Diseases  of  Animals  Act,  1894. 

15.  Information  to  be  given  as  to  Certain  Animals  or  Animals  in 

Contact  therewith. 

16.  Offences. 

17.  Extent. 

18.  Local  Authority  to  enforce  Order. 

19.  Commencement. 

20.  Short  Title. 

Schedule. 

205 


206  CLINICAL  BACTERIOLOGY  AND 


TUBERCULOSIS  ORDER  OF  1913. 

The  Board  of  Agriculture  and  Fisheries,  by  virtue  and 
in  exercise  of  the  powers  vested  in  them  under  the  Diseases 
of  Animals  Acts,  1894  to  1911,  and  of  every  other  power 
enabling  them  in  this  behalf,  do  order,  and  it  is  hereby 
ordered,  as  follows : 

Interpretation. 
1.   In  this  Order — 

"The  Board"  means   the   Board   of  Agriculture  and 

Fisheries  : 
"  Local  Authority  "  means  a  Local  Authority  for  the 

purposes  of  the  Act  of  1894 : 
"  The  Act  of  1894 "  means  the  Diseases  of  Animals 

Act,  1894 : 
"  Inspector  "  includes  Veterinary  Inspector  : 
"  Bovine  animal "  means  a   bull,  cow,  ox,  heifer,  or 

calf : 
"  Cow  "  includes  a  heifer  that  has  calved  : 
"Milk"  includes  cream    and   separated    or    skimmed 

milk. 

Other  terms  have,  where  the  context  so  permits,  the 
same  meaning  and  scope  as  in  the  Act  of  1894. 

Notice  of  Disease. 

2. — (1)  Every  person  having  in  his  possession  or  under 
his  charge 

(i.)  any  cow  which  is,  or  appears  to  be,  suffering  from 
tuberculosis  of  the  udder,  indurated  uddei*,  or 
other  chronic  disease  of  the  udder;  or 

(ii.)  any  bovine  animal  which  is,  or  appears  to  be, 
suffering  from  tuberculosis  with  emaciation 

shall  without  avoidable  delay  give  information  of  the  fact 
to  a  constable  of  the  police  force  for  the  area  wherein  the 
animal  is,  or  to  an  Inspector  of  the  Local  Authority,  and 
the  constable  or  Inspector  shall  transmit  the  information  to 
the  Local  Authority,  who,  if  not  themselves  the  Sanitary 
Authority,  shall  inform  that  Authority. 

(2)  The  person  in  possession  or  having  charge  of  the 
animal  shall  forthwith  take  such  steps  as  are  necessary  to 


VACCINE-THERAPY  207 

secure  compliance  with  Article  9  [Precautions  to  be  adopted 
with  Respect  to  Milk,  etc.)  and  Article  10  (Detention  and 
Isolation  of  Suspected  Animals). 

Notification  of  Disease  by   Veterinary  Surgeons. 

3. —  (1)  A  veterinary  surgeon  or  veterinary  practitioner 
who  in  his  private  practice  is  employed  to  examine  any 
animal,  and  is  of  opinion  that  the  animal,  if  a  cow,  is  suffer- 
ing from  tuberculosis  of  the  udder,  indurated  udder,  or 
other  chronic  disease  of  the  udder,  or,  if  a  bovine  animal, 
is  suffering  from  tuberculosis  with  emaciation,  shall  with 
all  practicable  speed  give  notice  of  the  existence  or  sus- 
pected existence  of  such  disease  to  an  Inspector  of  the 
Local  Authority,  who  shall  transmit  the  information  to  the 
Local  Authority,  who,  if  not  themselves  the  Sanitary 
Authority,  shall  inform  that  Authority. 

(2)  A  veterinary  surgeon  or  veterinary  practitioner  who 
under  and  in  accordance  with  this  Article  gives  notice  of 
the  existence  or  suspected  existence  of  disease  to  an 
Inspector  of  the  Local  Authority  shall  be  entitled  to  receive 
from  the  Local  Authority  a  fee  of  two  shillings  and  sixpence 
for  each  notification. 

(3)  Where  two  or  more  animals  are  examined  by  a 
veterinary  surgeon  or  veterinary  practitioner  on  the  same 
premises  and  at  the  same  time,  and  are  found  to  be  diseased, 
one  fee  only  shall  be  payable  to  him  under  this  Article  in 
respect  of  the  notification  of  the  existence  or  suspected 
existence  of  disease  in  such  animals. 


Inspection  and  Examination  of  Animals. 

4. —  (1)  Where  a  Local  Authority,  by  reason  of  informa- 
tion received  under  the  preceding  Articles  or  otherwise, 
have  reasonable  ground  for  supposing  that  on  any  premises 
in  their  District  there  is  a  cow  which  is  suffering  from 
chronic  disease  of  the  udder  or  giving  tuberculous  milk,  or 
a  bovine  animal  which  is  suffering  from  tuberculosis  with 
emaciation,  the  Local  Authority  shall  with  all  practicable 
speed  cause  such  veterinary  examination  of  the  bovine 
animals  on  such  premises  to  be  made  by  a  Veterinary 
Inspector  as  in  the  opinion  of  the  Local  Authority  is 
necessary  to  ascertain  whether  any  cow  thereon  is  suffering 
from  tuberculosis  of  the  udder  or  giving  tuberculous  milk, 


208  CLINICAL  BACTERIOLOGY  AND 

or  whether  any  bovine  animal  thereon  is  suffering  from 
tuberculosis  with  emaciation,  and  for  that  purpose  the 
Inspector  may,  with  the  previous  consent  in  writing  of  the 
owner  of  the  animal  or  of  his  agent,  but  not  otherwise, 
apply  the  tuberculin  test  to  any  cow  which  the  Inspector 
suspects  of  suffering  from  tuberculosis  of  the  udder,  or  of 
giving  tuberculous  milk,  or  to  any  bovine  animal  which  he 
suspects  of  suffering  from  tuberculosis  with  emaciation. 

(2)  For  the  purpose  of  such  examination,  a  Veterinary 
Inspector  may  at  all  reasonable  hours  enter  on  any  part  of 
the  premises  and  examine  any  bovine  animal  thereon,  and 
require  any  cow  to  be  milked  in  his  presence,  and  may  take 
samples  of  the  milk,  and  the  milk  from  any  particular  teat 
shall  if  he  so  require  be  kept  separate,  and  separate  samples 
thereof  shall  be  furnished. 

(3)  The  Inspector  may  also  take  samples  of  the  fseces  or 
urine  of  any  bovine  animal  on  the  premises,  or  of  any 
abnormal  discharge  from  any  bovine  animal  thereon. 

(4)  The  occupier  of  the  premises  and  the  persons  in  his 
employment  shall  render  such  reasonable  assistance  to  the 
Inspector  as  may  be  required  for  all  or  any  of  the  purposes 
of  this  Article,  and  any  person  refusing  such  assistance  shall 
be  deemed  guilty  of  an  offence  against  the  Act  of  1894. 

(5)  The  Inspector  shall  as  soon  as  possible  send  to  the 
Local  Authority  a  report  showing  the  result  of  his  inspec- 
tion and  examination  and  of  the  examination  of  any  sample 
taken  by  him.  The  Local  Authority,  if  not  themselves  the 
Sanitary  Authority,  shall  send  a  copy  of  the  report  to  that 
Authority. 

(6)  If  the  report  of  the  Inspector  as  to  any  animal  does 
not  show  that  it  is  suffering  from  tuberculosis  of  the  udder, 
or  giving  tuberculous  milk,  or  suffering  from  tuberculosis 
with  emaciation,  the  Local  Authority  shall  forthwith  give 
notice  in  writing  to  the  owner  or  person  in  charge  thereof 
that  the  provisions  of  this  Order  relating  to  precautions  to 
be  adopted  with  respect  to  milk  and  detention  and  isolation 
of  suspected  animals  have  ceased  to  apply  to  the  animal. 

Slaughter  of  Diseased  Animals. 

5. — (1)  Where  a  Local  Authority  are  satisfied  by  the 
report  of  the  Inspector  that  in  their  District  there  is  a  cow 
which  is  suffering  from  tuberculosis  of  the  udder,  or  giving 
tuberculous  milk,   or  a  bovine  animal   which  is   suffering 


VACCINE-THERAPY  209 

from  tuberculosis  with  emaciation,  the  Local  Authority  shall 
with  all  practicable  speed  give  notice  in  writing  (in  the 
Form  set  forth  in  the  Schedule  hereto  or  to  the  like  effect) 
to  the  owner  or  person  in  charge  of  the  animal  and  also  to 
the  Board,  and  cause  the  animal  to  be  slaughtered  j  provided 
that  if,  before  the  slaughter  is  carried  out,  the  owner  of  the 
animal,  or  any  person  on  his  behalf,  gives  notice  in  writing 
to  the  Local  Authority,  or  to  their  Inspector  or  other  officer 
lirected  to  carry  out  such  slaughter,  that  the  owner  objects 
to  the  animal  being  slaughtered  under  the  provisions  of  this 
Order,  it  shall  not  be  lawful  for  the  Local  Authority  to 
cause  the  animal  to  be  slaughtered  without  the  special 
authority  of  the  Board  first  obtained;  provided  also  that 
this  special  authority  shall  not  be  given  in  the  case  of  any 
animal  valued  under  this  Order  at  more  than  thirty  pounds, 
if  and  so  long  as  the  animal  is  detained  and  isolated,  and 
the  milk  (if  any)  is  dealt  with  in  accordance  with  the 
provisions  of  this  Order. 

(2)  If  the  value  of  an  animal  proposed  to  be  slaughtered, 
as  agreed  or  certified  under  this  Order,  exceeds  thirty 
pounds,  the  Local  Authority  shall  not  proceed  with  its 
slaughter  unless  so  directed  by  the  Board. 

Valuation  for  Compensation. 

6. —  (1)  Before  the  slaughter  of  an  animal  the  Local 
Authority  shall  either  agree  in  writing  with  the  owner  of 
the  animal  the  value  thereof  in  its  condition  at  the  time  of 
valuation,  or  if  they  shall  fail  so  to  agree  shall  cause  such 
value  to  be  ascertained  by  a  valuer  appointed  by  them  or 
appointed  on  the  application  of  the  Local  Authority  by  the 
Board,  but  paid  by  the  Local  Authority,  and  such  valuer 
shall  give  to  the  Local  Authority  and  to  the  owner  a 
certificate  in  writing  of  the  said  value. 

(2)  In  ascertaining  the  value  of  an  animal,  regard  shall 
be  had  to  any  Act,  Order,  or  Regulation  dealing  with  the 
sale  or  use  of  milk,  milk  products,  or  carcases  for  human 
food. 

(3)  The  value  shall  be  ascertained  both  on  the  basis  of 
the  certificate  of  examination  hereinafter  required  showing 
that  the  animal  was  suffering  from  tuberculosis,  and  alsoon 
the  basis  of  its  not  showing  that  the  animal  was  suffering 
from  tuberculosis,  and  the  amount  to  be  paid  for  com- 
pensation shall  depend  on  such  certificate  accordingly. 


210  CLINICAL  BACTERIOLOGY  AND 

Post-Mortem  Examination  of  Slaughtered  Animals. 

7. —  (1)  In  the  case  of  every  animal  slaughtered  under 
this  Order,  the  Local  Authority  shall  cause  the  carcase,  at 
the  time  of  slaughter  or  as  soon  as  practicable  thereafter, 
to  be  examined  by  a  Veterinary  Inspector  of  the  Local 
Authority,  or  (if  so  required  by  the  owner  or  person  in 
charge  of  the  animal  before  it  is  slaughtered)  by  some  other 
veterinary  surgeon,  who,  failing  agreement  between  the 
Local  Authority  and  such  owner  or  person,  shall  be  nomi- 
nated by  the  Board,  but  paid  by  the  Local  Authority. 

(2)  The  Veterinary  Inspector  or  other  veterinary  surgeon 
shall  at  the  conclusion  of  his  examination  give  to  the  Local 
Authority  and  to  the  owner  of  the  animal  a  certificate  of 
the  result  of  the  examination  in  the  Form  set  forth  in  the 
Schedule  hereto  or  to  the  like  effect. 

Compensation. 

8. — (1)  If  the  Local  Authority  fail  to  carry  out  the 
examination  required  by  the  preceding  Article,  or  if  the 
certificate  of  such  examination  does  not  show  that  the 
animal  was  suffering  from  tuberculosis,  the  Local  Authority 
shall,  by  way  of  compensation,  pay  to  the  owner  thereof 
a  sum  equal  to  the  value  of  the  animal  as  agreed  or 
certified  in  manner  aforesaid,  and  a  further  sum  of  twenty 
shillings. 

(2)  If  the  certificate  of  the  examination  shows  that  the 
animal  was  suffering  from  tuberculosis  (not  being  advanced 
tuberculosis),  the  Local  Authority  shall,  by  way  of  com- 
pensation, pay  to  the  owner  a  sum  equal  to  three-fourths 
of  the  value  of  the  animal  as  agreed  or  certified  in  manner 
aforesaid,  after  deducting  from  such  sum  one-half  of  their 
reasonable  costs  of  any  valuation  of  the  animal  by  a  valuer 
appointed  by  the  Board,  and  of  any  examination  of  its 
carcase  by  a  veterinary  surgeon  other  than  the  Veterinary 
Inspector. 

(3)  If  the  certificate  of  the  examination  shows  that  the 
animal  was  suffering  from  advanced  tuberculosis,  the  Local 
Authority  shall,  by  way  of  compensation,  pajr  to  the  owner 
a  sum  equal  to  one-fourth  of  the  value  of  the  animal,  as 
agreed  or  certified  in  manner  aforesaid,  or  the  sum  of  thirty 
shillings,  whichever  sum  is  the  greater,  after  deducting 
from  such  sum  one-half  of  their  costs  of  valuation  and 
examination  as  in  the  preceding  case. 


VACCINE-THERAPY  211 

(4)  For  the  purposes  of  this  Order  an  animal  slaughtered 
under  this  Order  shall  be  deemed  to  have  been  suffering 
from  advanced  tuberculosis 

(a)  when  there  is  miliary  tuberculosis  of  both  lungs; 

(b)  when  tuberculous  lesions  are  present  on  the  pleura 

and  peritoneum; 

(c)  when  tuberculous  lesions  are  present  in  the  mus- 

cular system,  or  in  the  lymphatic  glands  embedded 
in  or  between  the  muscles ;  or 

(d)  when  the  carcase  is  emaciated  and   tuberculous 

lesions  are  present. 

Precautions  to  be  adopted  with  Respect  to  Milk,  etc. 

9. — (1)  The  milk  produced  by  any  cow  which  is,  or 
appears  to  be,  suffering  from  chronic  disease  of  the  udder 
or  tuberculosis  with  emaciation,  shall  not  be  mixed  with 
other  milk  until  the  cow  has  been  examined  by  a  Veterinary 
Inspector  in  accordance  with  the  provisions  of  this  Order, 
and  until  the  owner  or  person  in  charge  thereof  has  been 
notified  that  this  Article  has  ceased  to  apply  to  the  cow ; 
and  all  milk  affected  by  this  Article  shall  forthwith  be 
boiled  or  otherwise  sterilized,  and  any  utensil  in  which  such 
milk  is  placed  before  being  so  treated  shall  be  thoroughly 
cleansed  with  boiling  water  before  any  other  milk  is  placed 
therein. 

(2)  A  Local  Authority,  or  a  Veterinary  Inspector  on 
their  behalf,  may  by  written  notice  apply  the  restrictions 
imposed  by  this  Article  to  the  milk  produced  by  any  cow 
specified  in  the  notice  which  is  suspected  of  giving  tuber- 
culous milk  and  is  being  examined  under  this  Order,  and 
such  restrictions  shall  apply  accordingly. 

Detention  and  Isolation  of  Suspected  Animals. 

10. —  (1)  Every  person  having  in  his  possession  or  under 
his  charge  any  cow  which  is,  or  appears  to  be,  suffering 
from  chronic  disease  of  the  udder,  or  any  bovine  animal 
which  is,  or  appears  to  be,  suffering  from  tuberculosis  with 
emaciation,  shall  keep  the  animal  isolated  as  far  as  prac- 
ticable from  other  bovine  animals,  and  also  keep  the  animal 
in  his  possession  or  under  his  charge,  until  the  animal  has 
been  examined  by  a  Veterinary  Inspector  in  accordance 
with  the  provisions  of  this  Order  and  the  owner  or  person 


212  CLINICAL  BACTERIOLOGY  AND 

in  charge  thereof  has  been  notified  that  this  Article  has 
ceased  to  apply  to  the  animal ;  provided  that  the  animal 
may  at  any  time  be  slaughtered  by  the  owner  or  person  in 
charge. 

(2)  A  Local  Authority,  or  a  Veterinary  Inspector  on 
their  behalf,  may  by  written  notice  apply  this  Article  to 
any  bovine  animal  specified  in  the  notice  which  is  being 
examined  under  this  Order,  and  such  Article  shall  apply 
accordingly. 

Suspected  Animals  in  Markets,  Fairs,  and  Sales. 

11. — (1)  A  Veterinary  Inspector  of  a  Local  Authority 
may  by  notice  served  on  the  owner  or  person  in  charge  of 
a  bovine  animal  exposed  in  a  market,  fairground,  or  sale- 
yard,  which  appears  to  him  to  be 

(i.)   suffering  from  tuberculosis  of  the  udder,  indurated 
udder,  or  other  chronic  disease  of  the  udder ;  or 
(ii.)   suffering  from  tuberculosis  with  emaciation, 

require  the  animal  to  be  removed  from  the  market,  fair- 
ground, or  saleyard,  to  the  premises  from  which  it  was 
brought  thereto,  or,  if  the  owner  or  person  in  charge  so 
desires,  to  any  other  suitable  premises,  to  be  specified  in 
the  notice,  and  thereupon  the  animal  shall  forthwith  be 
moved  by  the  owner  or  person  in  charge  to  those  premises 
for  the  purpose  of  examination  under  the  foregoing  pro- 
visions of  this  Order. 

(2)  Where  the  premises  to  which  the  animal  is  required 
under  this  Article  to  be  moved  are  not  in  the  same  District 
as  the  market,  fairground,  or  saleyard,  the  Inspector  serving 
the  notice  shall  forthwith  send  a  copy  of  the  notice  to  the 
Local  Authority  of  the  District  in  which  the  first-mentioned 
premises  are  situate. 

Cleansing  and  Disinfection. 

12.  The  occupier  of  any  premises  on  which  there  has 
been  a  cow  suffering  from  tuberculosis  of  the  udder  or 
giving  tuberculous  milk,  or  a  bovine  animal  suffering  from 
tuberculosis  with  emaciation,  shall,  if  so  required  in  writing 
by  an  Inspector  of  the  Local  Authority,  cleanse  and  dis- 
infect at  his  own  expense,  and  to  the  satisfaction  of  the 
Inspector,  that  part  of  any  shed  or  other  erection  in  which 
the  animal  has  recently  been  placed  or  kept. 


VACCINE-THERAPY  213 

Reports  to  the  Board. 

13.  Every  Local  Authority  and  their  Inspectors  and 
officers  shall  send  and  give  to  the  Board  such  reports, 
returns,  and  information,  as  to  their  proceedings  under  this 
Order,  as  the  Board  require. 

Extension  of  Certain  Sections  of  Diseases  of  Animals 
Act,  1894. 

14.  Tuberculosis  shall  be  a  disease  for  the  purposes  of 
the  following  sections  of  the  Act  of  1894  (namely)  : 

Sections  nineteen  and  twenty  {Slaughter  in  Disease  and 
Compensation  generally) ; 

Section  forty-three  (Police)  ; 

Section  forty-four  (General  Administrative  Provisions)  ; 
and  also  for  the  purposes  of  all  other  sections  of  the  said 
Act  containing  provisions  relative  to  or  consequent  on  the 
provisions  of  those  sections  and  this  Order,  including  such 
sections  as  relate  to  offences  and  legal  proceedings. 

Information  to  be  given  as  to  Certain  Animals  or  Animals 
in  Contact  therewith. 

15.  Article  36  of  the  Animals  (Transit  and  General) 
Order  of  1912  (Information  to  be  given  as  to  Diseased  or 
Suspected  Animals  or  Animals  in  Contact  therewith)  shall 
apply  to 

(i.)  any  cow  which  is,  or  is  suspected  of,  suffering  from 
tuberculosis  of  the  udder  or  giving  tuberculous 
milk ;  and 

(ii.)  any  bovine  animal  which  is,  or  is  suspected  of, 
suffering  from  tuberculosis  with  emaciation. 

Offences. 

16.  Every  person  who — 

(i.)   fails  to   give   the  notice   required  by  Article  2  or 

Article  3  of  this  Order ;  or 
(ii.)   fails  to  comply  with  any  provision  of  this  Order 

relating  to'  precautions  to  be  adopted  with  re- 


214  CLINICAL  BACTERIOLOGY  AND 

spect  to  milk  or  relating  to  detention  and  isola- 
tion of  animals;  or 
(iii.)  fails  to  comply  with  any  notice  directing  removal  of 
an  animal  from  a  market,  fairground,  or  sale- 
yard;  or 

(iv.)  fails  to  cleanse  or  disinfect  any  erection  which 
under  this  Order  he  is  required  to  cleanse  or  dis- 
infect, 

shall,  according  to  and  in  respect  of  his  own  acts  and 
defaults,  be  deemed  guilty  of  an  offence  against  the  Act 
of  1894.  *      '  * 

Extent. 

17.  This  order  extends  to  England  and  Wales  and 
Scotland. 

Local  Authority  to  enforce  Order. 

18.  The  provisions  of  this  Order,  except  where  it  is 
otherwise  provided,  shall  be  executed  and  enforced  by  the 
Local  Authority. 

Commencement. 

19.  This  Order  shall  come  into  operation  on  the  first 
day  of  May,  nineteen  hundred  and  thirteen. 


Short  Title. 

20.    This  Order  may  be  cited  as  the  Tuberculosis  Order 
of  1913. 

In  witness  whereof  the  Board  of  Agriculture  and 
Fisheries  have  hereunto  set  their  Official  Seal  this 
thirteenth  day  of  February,  nineteen  hundred  and 
thirteen. 

Sydney  Olivier, 

Secretary. 


VACCINE-THERAPY  215 


SCHEDULE. 


Forms. 

TUBERCULOSIS  ORDER  OF  1913. 
Form  of  Notice  of  Intended  Slaughter. 
(Article  5.) 
To  of 

The  Local  Authority  for  the  county  [borough  or  burgh]  of 
hereby  give  notice  that  they  are  satisfied  that  [insert  description  of 
animal]  which  is  now  kept  at  [insert  description  of  premises  where  it 
is  kept,  stating  parish]  is 

*(a)  suffering  from  tuberculosis  of  the  udder  ; 

*(b)  giving  tuberculous  rnilk  ; 

*(c)  is  suffering  from  tuberculosis  with  emaciation, 

and  that  they  propose  with  all   convenient  speed   to   slaughter   the 
animal. 

*  Strike  out  the  part  which  is  inapplicable. 

(Signed) 

By  direction  of  the  Local  Authority. 

Dated  191     . 

Note. — If  the  owner  of  the  animal,  or  any  person  on  his  behalf 
before  the  slaughter  is  carried  out,  gives  notice  in  writing  to  the  Local 
Authority,  or  to  their  Inspector  or  other  officer  directed  to  carry  out 
the  slaughter,  that  the  owner  objects  to  the  animal  being  slaughtered, 
it  may  not  be  slaughtered  without  the  special  authority  of  the  Board 
first  obtained. 

The  compensation  payable  by  the  Local  Authority  is  regulated  by 
the  Order. 


TUBERCULOSIS   ORDER   OF   1913. 

Form  of  Certificate  of  Result  of  Post-Mortem  Examination. 

(Article  7.) 

I,  A.B.,  a  Veterinary  Inspector  of  the  Local  Authority  for  the  county 
[borough  or  burgh]  of  [or  a  veterinary  surgeon  acting 

under  the  Tuberculosis  Order  of  1913],  do  hereby  certify  that  my 
examination  of  the  carcase  of  [here  describe  the  animal  slaughtered], 
which  was  caused  to  be  slaughtered  by  the  Local  Authority  for  the 


216  CLINICAL  BACTERIOLOGY 

county  [borough  or  burgh]  of  on  the 

day  of  191     ,  and  which  animal  belonged  to 

of  ,  does  not  show  that  the  animal  was  affected  with 

tuberculosis  [or  shows  that  the  animal  was  affected  with  tuberculosis 
(not  being  advanced  tuberculosis  within  the  meaning  of  the  Tubercu- 
losis Order  of  1913)]  [or  shows  that  the  animal  was  suffering  from 
advanced  tuberculosis  within  the  meaning  of  the  Tuberculosis  Order 
of  1913]. 

(Signed)  A.B. 

Dated  191    . 


BIBLIOGRAPHY 

Allen  :  Vaccine  Therapy. 

Bowhill  :  i  Manual  of  Bacteriological  Technique  and  Special  Bac- 
teriology. 

Buchanan  :  Veterinary  Bacteriology. 

Emery  :  Clinical  Bacteriology  and  Hematology. 

Friedberger  and  Frohner  :  Veterinary  Pathology. 

Hewlett  :  Manual  of  Bacteriology. 

Hutyra  and  Marek  :  Special  Pathology  and  Therapeutics  of  the 
Diseases  of  Domesticated  Animals. 

Jowett  :  Notes  on  Blood-Serum  Therapy. 

Law  :  Veterinary  Medicine. 

Wright  :  Technique  of  the  Teat  and  Capillary  Glass  Tube. 


217 


INDEX 


Abscess  : 

bacteria,  91 
bacteriology  of,  91,  92 
treatment  of,  92 
Acne: 

bacteria  in,  43,  101 
cultivation  of   acne  bacillus, 
101,  102 
Actinomycosis,    44,   45,   91,    142, 

165,  166 
Agglutinins,  59 
Alexins,  56 
Amboceptor,  61,  62 
Anaerobic  cultivation,  23 

by  Friinkel's  method,  24 
by  the  glucose  method,  23 
by    the    pyrogallic    acid 
method,  24 
Anal  fistula,  100,  101 
Anaphylaxis,  89,  182 
Anthrax,  125,  133 

serum  treatment  by  Deutsch, 
Legge,     Mendez,     Sclaro, 
Sobernheim,  134,  135 
Vaccine  No.  1,  133,  134 
Vaccine  No.  2,  134 
vaccine   treatment   (Pasteur) 
in,  133,  134 
Anti-anaphylaxis,  182 
Antibodies,  56 

non-specific,  56 
specific,  56 

in  tuberculosis,  199,  200 
Antiformin,  176 
Antistreptococcic  serum,  149 
in  endometritis,  124 
in  influenza,  155 
in  poll-evil,  96 
in  pyaemic  arthritis,  110 
in    septicemic    arthritis, 

112 
in  strangles,  149 


Antitetanin,  140 
Antitoxins,  56 
Arthritis,  105 

pyaemic,   in   foals,   106,    107, 

108,  109,  110 
septicamiic,     110,    111,    112, 

113 
traumatic,  105,  106 
Autogenous  vaccine,  dose  of,  85, 


Bacilli,   differential   diagnosis   of 

38-46 
Bacillus : 

acne,  43,  91 
anthracis,  43,  164 

symptomatica,   40,    125, 
130 
bronchosepticus,  40,  159, 162, 

163 
coli  communis,  40,  100,  109, 
110,  114,  115, 116,  117, 118, 
119, 124,  125,  126,  127,  142 
diphtherias,  44 
mallei,  45,  157 
cedematis  maligni,  39,  129 
of  Friedlander,  45 
pyocyaneus,  39,  91,  100,  109, 

110,  115,  119 
suipestifer,  188,  189 
suisepticus,  188,  189 
tetani,  38,  138,  139 
tuberculosis,  43,  91,  175,  177, 
178 
Bacteriolysis,  61 
Balances,  1 
Blackquarter,  125,  130 

Arloing's   double   method   of 

vaccination,  132 
inoculation  in,  132,  133 
Kitt's  single  method  of  vac- 
cination, 131 


218 


INDEX 


219 


Blood  capsules  (Wright's),  51,  52 
collection  of,  51,  52 
film  stains,  33 

how  made,  53,  54 
Jenner's,  33 
Leishman's,  33 
protective  elements  of,  55 
Bovine  tuberculosis,  142-167 

collecting  materia  morbi 

in,  175 
examining  materia  morbi 

in,  176 
of  the  alimentary  system, 

171,  172 

of  the  generative  system, 

172,  173 

of  the  lymphatic  system, 

173 
of  the  mammary  system, 

174 
of  the   nervous   system, 

172 
of  the  respiratory  system, 

169,  170,  171 
post-mortem   lesions   in, 

184 
staining  the  bacillus  in, 
177 
Pappenheim's 

method,  178 
Ziehl  -  Neelsen's 
method,  177-178 
staphylococci  in,  167 
streptococci  in,  168 
tests  in : 

biological,  182,  183 
tuberculin,    use    of, 
and  how  made, 
179 
cutaneous,     181, 

182 
ophthalmic,  181 
subcutaneous, 
179,  180 
Bronchitis,  142,  143,  144, 145, 146 

Calmette's    ophthalmo  -  reaction, 

181 
Canine  distemper,  142,  158,  159, 
160,  161,  162, 163, 164, 
165 
agglutination  test  in,  163 
protective    inoculation 
against,  163,  164,  165 


Canine   distemper,    researches   of 
Carre,    Copeman,   Ferry,    Lijj- 
nieres,    Piorkowski,  Rale,  Val- 
lerio,    M'Gowan,    Torry,     and 
Hake,  in,  158, 159 
Capillary  pipettes,  47.  48,  49 
Capsule-staining,  31 
Centrifugalizing  blood,  13 
milk,  14 
urine,  13,  14 
Centrifuge,  11 
electric,  12 
hand,  12 
water,  12 
Cocci : 

catarrhalis,  38,  142,  144 
differential   diagnosis   of,  36, 

37,38 
diplococci,  37,  127,  147,  151 
pneumococci,  37,  142,  144 
staphylococci,  36.  91,  92,  94, 
95,  100,  101,  102,  105,  109, 
110,  113,  114,  119,  125, 127, 
142,  144,  151,  165,  168 
streptococci,  36,  91,  100,  101, 
105, 108, 110, 112, 113, 114, 
118, 119,  124,  125, 127,  142, 
144,  148,  168 
tetrageni,  37,  109,  142 
Cocco  bacillus,  147 
Cornet  forceps,  27 
Cultivation  of  bacteria,  20,  21 
anaerobic,  23,  24 
plate— Petri  dish,  21,  22 
Culture  media,  15 
Cystitis,  120 

Dose  of  autogenous  vaccine,  85, 
86 
of  vaccine,  83 
Diarrhoea,  114 

Distemper.    See  Canine  distemper 
Dysentery,  114 

Ehrlich  side-chain  theory,  57,  58 
Emery  paper,  4,  53 
Endometritis,  114,  121 

antistreptococcal    serum    in, 
124 

collecting  materia  morbi  in, 
122 
Endotoxins,  57 

Equine  influenza.     See  Influenza 
Exotoxins,  57 


220 


INDEX 


Films  : 

fixing,  27 

by  alcohol,  27 

by   corrosive   sublimate, 

27 
by  formalin,  28 
by  heat,  27 
Filtf-rable  virus  group,  89 
Fistula  of  the  poll,  92,  98 

of  the  praesternum,  99,  100 
of  the  rectum  and  anus,  100, 

101 
of  the  withers,  92,  98 
Flagella-staining,  31 
Flasks,  3 
Follicular  mange,  102,  103 

Glanders,  142,  157 

bacillus  of,  45 

mallein  reaction  in,  157 
test  in,  158 

Widal's  agglutination  test  in, 
157 
Glass-cutting  knife,  47 

tubing,  47 
Glasswork  requisites,  47 
Gram-negative  bacteria,  29 

-positive  bacteria,  29 

Hanging-drop,  preparation  of,  34, 

35 
Haptophore,  58,  139 
Hot-air  oven,  3 
Hydrophobia,  136,  137,  138 
Hyper-immune,  191 

Identification  of  bacteria,  34 
Illumination,  6 

dark-ground,  4 
In.'nmnization  by  vaccine,  80 

phases  following — (a)  Nega- 
tive,   (6)   positive,    (c)    in- 
creased resistance,  81 
Incubator,  7,  8,  9 
Influenza,    142,    153,    154,    155, 

156 
Inoculating  medium  tubes,  20 
Involution  forms  of  bacteria,  84 

Jennets  blood-stain,  33 
Johne's  disease,  116 

Knife,  glass-cutting,  47 
Koch's  sterilizer,  11 


Laboratory,  1 

accessories,  1,  2,  3 

requisites  (improvised),  3,  4 
Leishman's  stain,  33 
Lenses,  immersion,  4,  5 

microscopical,  4 
Lockjaw.     See  Tetanus 
Lysol,  4 

Malignant  oedema,  125,  129,  130 
protective  serum  in,  129 
Mallein,  157 

in  diagnosis,  157 
preparation  of,  157 
Mange  in  the  dog,  102 
Measures  and  weights,  201 
Media  culture,  14 

agar-agar,  16 
blood-serum,  16 
broth,  15 
gelatin,  16 
potato,  17 
Medium  tubes,  18 
sloped,  18 
stab,  21 
filling  test-tubes  with,  17 
Micrococcus  catarrhalis,  38 

tetragenus,  37,  109 
Microscope,  4 

the  use  of,  6,  7 
Milk: 

centrifugalizing,  14 

how    to    remove,    from    the 

udder,  176 
in  tuberculosis  of  the  udder, 
174 
Minimum    dose   of    vaccine,  85, 
86 

Navel  ill,  106,  107,  108,  109,  110 
Negre  bodies,  137 
Nephritis.     See  Pyelonephritis 
Non-specific  antibodies,  56 

Objectives,  4 

(Edema,  malignant,  125 

Oil  immersion  lens,  4,  6 

Ophthalmo  -  reaction  in  tubercu- 
losis, 181 

Opsonic  index : 

determination  of,  63,  64,  65, 

66,  67,  68 
in  influenza,  155 
in  mange  follicules,  103 


INDEX 


221 


Pappenheim's  solution  for  tubercle, 

178 
Pasteur  vaccine  in  anthrax,  133, 

134 
Pasteur's  treatment  of  rabies,  137 
Peritonitis,  114,  117 

B.  coli  in,  117,  118 
Petri  dishes,  22 
Pfeiffer's  reaction,  161,  162 
Phagocytosis,  68 
Pipettes,  47,  48,  49 
Pitfield's  method,  31 
Platinum  needles,  20 
Pneumococci,  37 
Pneumonia,  142,  146,  147 

cocco  bacillus  in,  147 
Poll-evil,  92,  93,  94,  95,  96,  97,  98, 

99 
Polyvalent  vaccine,  71 
Post-mortem  : 

swine  fever,  189 

tuberculosis,  184 
Potato.     See  Media 
Prsesternal  fistula,  99,  100 
Precipitation  test,  61 
Purpura  haemorrhagica,  150,  151 
Lugal's  solution  in,  153 
Pyaemia,  125,  128,  129 
Pyaernic    arthritis    in    foals,    105, 

107,  108,  109,  110 
Pyelonephritis,  118,  119,  120 

drugs  in,  120 

examination  of  urine  in,  119, 
120 

uraemia  in,  120 

Quarter-evil,  125,  130 

Rabies,  136,  137,  138 

Hogge's  treatment  in,  138 

Pasteur  on,  138 
Ray  fungus  in  actinomycosis,  165 
Receptors,  58 
Retention  of  placenta,  121 
Roaring,  prophylactic  sero-vaccine 
in,  157,  197,  198,  199 

Sealing  glass  tube,  51 

test-tube,  49 
Septicaemia,  125 
Septicaemic  arthritis,  110,  111,  112, 

113 
Septic  infection,  125,  126 

intoxication,  125,  126 


Serum  : 

antibacterial,  87 
disease,  88 

anaphylaxis  in,  89 
mode  of  preparation,  87 
therapy  abroad,  194,  195, 196 
Side-chain  theory,  58 
Sink,  1 
Slides,  glass,  53 

concave  face  of,  53 
convex  face  of,  53 
how   to    remove    grease 
from,  53 
Spore-staining,  32 
Stage,  microscopical,  6 
Staining  methods,  25 
Stains,  28 

aniline  gentian  violet,  29 
carbol fuchsin,  30 

thionin,  30 
eosin,  31 

methylene  blue,  30 
neutral  red,  31 
Standardization  of  vaccine,  73, 74, 

75,  76 
Staphylococci,  36,  91,  94,  95,  100, 
101,  103,  105,  108, 109,  110,  113 
Steam  sterilizer,  11 
Sterilizers,  9 

dry  heat,  10 
moist  heat,  10 
Strangles,  142,  148 

antistreptococcal    serum    in, 
149 
Streptococci,  36,  91, 100,  101,  105, 

108,  110,  112,  113 
Swine  fever,   142,   186,  187,  188, 
189,  190,  191,  192,  193 
bacillus     suipestifer     in, 
188,  189 
suisepticus   in,    188, 
189 
method  of  making  serum, 
191,  192 
of  preserving  serum, 
192 
post-mortem   lesions  in, 

189,  190 
serum  -  therapy   in,    190, 

197 
symptoms  of  abdominal 
form  in,  187 
of  pulmonary  form 
in,  187 


222 


INDEX 


Swine   fever,  symptoms    of    sep- 
ticsemic  form  in,  186 
ultra-visible  virus  in,  186 
Symptomatic  anthrax.   See  Black- 
quarter 
Syringes : 

serum,  77,  78 
vaccine,  77,  78 

Tetanus,  136 

bacillus,  38,  39 
Behring's  serum  in,  140 
Theobald  Smith's  phenomena,  89 
Thermolable,  58 
Thermostable,  58 
Toxins,  57,  58 
Toxoids,  58 

Tubercle  bacillus,  43,  44,  176,  177, 
178 
in  expectorate,  175 
in  faeces,  176 
in  milk,  176 
in  tissues,  175,  176 
in  urine,  176 
staining,  177,  178 
Tuberculin : 

cutaneous  test  with,  181,  182 
how  made,  179 
ophthalmic  test  with,  181 
reaction  with,  180 
subcutaneous  test  with,  179, 

180 
treatment  with,  168 
Tuberculosis,  bovine : 

biological   diagnosis   of,  182, 
183,  184 


Tuberculosis,  bovine — continued : 
clinical  diagnosis  of,  167, 168, 

169,  170,  171,  172,  173,  174 
macroscopical    diagnosis    of, 

184,  185 
microscopical    diagnosis    of, 

177,  178 

Ulcer,  104 

Ultra-visible  virus  group,  89 
Urine,  bacteriological  investigation 
of,  109,  120 

Vaccine : 

age  of  cultures  for,  84 
autogenous,  71 
curative,  71 
dose  of,  83,  85 
immunization  by,  80 
mode  of  preparation  of,  69 
monovalent,  71 
polyvalent,  71 
prophylactic,  71 
standardization    of,    73,    74, 
75,  76 

Weights  and  measures,  201 
White  scour  in  calves,  115,  116 
Widal's  agglutination  test,  59,  60 
Withers,  fistula  of,  92,  93,  94,  95, 

96,  97,  98,  99 
Wright's  blood  capsules,  51,  52 
spreader,  25,  26 


Ziehl-Neelsen's     stain, 
178 


30,    177. 


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LAVERAN  &  MESNIL'S  Trypanosomes  and  Trypanosom- 
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with  coloured  Plate  and  81  Illustrations.     Price  21s.  net,  postage  6d. 

LAW  S  Text-Book  of  Veterinary  Medicine. 

In  Five  Volumes.  Vol.  I.:  General  Pathology;  Diseases  of  the  Res- 
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MALKMUS'  Outlines  of  Clinical  Diagnosis  of  the  Internal 
Diseases  of  Domestic  Animals.    Fourth  Edition.    Pp.  259,  with 

57  Illustrations.     Price  12s.  6d.  net,  postage  5d. 

MERILLAT'S   Animal   Dentistry  and   Diseases   of   the 

Mouth.     Pp.  '262,  with  160  Illustrations.     Price  12s.  6d.  net,  postage  4d. 

MERILLAT'S  Principles  of  Veterinary  Surgery. 

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MULLER'S  Diseases  of  the  Dog-  and  their  Treatment. 

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plain  and  coloured  Illustrations.     Price  25s.  net,  postage  7d. 

NEUMANN'S  Treatise  on  the  Parasites  and  Parasitic 

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Illustrations.     Price  21s.  net,  postage  6d. 

OSTERTAG'S  Handbook  of  Meat  Inspection. 

Fourth  Edition.  Translated  by  E  V.  Wilcox,  M.A.  Pp.  xxxvi  +  886, 
with  coloured  Plate  and  260  Illustrations.     Price  31s.  6d.  net,  postage  7d. 

REEKS'  Common  Colics  of  the  Horse. 

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postage  4d. 

REEKS'  Diseases  of  the  Horse's  Foot. 

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ROBERTSON'S  Meat  and  Food  Inspection. 

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SCOTT'S  Clinical  Bacteriology  and  Vaccine  Therapy  for 

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trations in  the  Text.     Price  7s.  6d.  net,  postage  5d. 

SMITH'S  Manual  of  Veterinary  Physiology. 

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•will  afiord  pleasure  for  leisure  hours,  and  can  be  read  with  marked  profit  by 
the  practitioner.  After  its  perusal  he  will  recognize  that  it  is  the  work  of  one 
who  is  not  o  ly  an  accomplished  scientist,  but  also  a  skilled  clinician — a  com- 
bination which,  we  may  remark,  is  very  seldom  met  with." — Veterinary  News. 

Third  Edition.  Pp.  xvi  +  715,  with  coloured  Plate  and  160  Illustrations. 
Price  15s.  net,  postage  5d. 

SMITH'S  Manual  of  Veterinary  Hygiene. 

Third  Edition.  Pp.  xx  +  1036,  with  225  Illustrations.  Price  15s.  net, 
postage  7d. 

SMYTHE'S  Veterinary  Parasitology. 

Owing  to  the  increased  attention  paid  to  the  study  of  parasitology,  there 
has  arisen  a  need  in  our  literature  of  a  text-book  dealing  with  the  more 
common  parasites  of  the  domesticated  animals.  It  has  been  the  aim  of  the 
author  to  fill  the  gap  with  a  volume  which,  while  covering  the  course  laid 
down  for  the  student,  shall,  at  the  same  time,  be  of  practical  value  to  the 
veterinarian  as  a  book  of  reference. 

Pp.  xiv  +  130,  with  29  Illustrations.     Price  4s.  net,  postage  4d. 


An  Abridged  List  of  Works. 


THOMPSON'S    Elementary    Lectures     on     Veterinary 
Science. 

A  particular  fea'ure  of  this  new  edition  is,  that  the  whole  of  the  illustrations 
have  been  redrawn.  For  its  particular  purpose  this  book  is  the  best  of  ita 
kind  iu  existence.  It  should  form  p;rt  of  the  equipment  of  every  veterinary 
student. 

Fourth  Edition.  Pp.  xvi  +  544,  with  54  Plates  and  17  Illustrations  in 
the  Text.     Price  10s.  6d.  net,  postage  5d. 

TWORT  &  INGRAM'S  Johne's  Disease. 

In  this  book  an  attempt  has  been  made  to  summarize  existing  knowledge  on 
the  subject,  and  the  authors  have  brought  before  English  readers  the  investiga- 
tions of  Continental  workers,  which  have  not  been  generally  available  to 
veterinary  surgeons  in  this  country.  The  volume  als  >  includes  their  own 
investigations,  which  have  recently  been  carried  out  at  the  Brown  Institution. 

Pp.  xii  +  180,  with  9  Plates.     Price  6s.  net,  postage  4d. 

"WILLIAMS'    Principles    and    Practice   of    Veterinary 

Medicine.  Ninth  Edition.  Revised  by  W.  Owen  Williams, 
F.R.C.V.S.,  and  F.  S.  H.  Baldrey,  Major  I.CV.D.,  F.R.C.V.S., 
D.V.H.  Ltv.  Pp.  xviii  +  98c>,  with  25  Plates,  mostly  coloured,  and 
90  Illustrations  in  the  text.     Price  25s.  net,  postage  7d. 

WINSLOWS  Veterinary  Materia  Medica  and  Therapeutics. 

(According  to  the  British  and  United  States  Pharmacopoeias.)  Fifth 
Edition.     Pp.  vii  +  857.     Price  25s.  net,  postage  6d. 

WOODRUFF'S  Economies  of  Feeding"  Horses. 

"  The  object  of  the  work  is  to  explain  the  principles  of  feeding,  and  to  show 
how  those  principles  cm  be  put  into  practice  with  economic  advantage.  It  is 
written  for  the  use  of  veterinary  suiyeons,  for  horse  owners  and  managers  of 
studs,  to  all  of  whom  the  subject  is  of  the  greatest  importance."— Veterinary 
Journal. 


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TROPICAL  VETERINARY  BULLETIN. 

Published  Quarterly  for  the  Tropical  Diseases  Bureau,  Imperial  Institute, 
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VETERINARY  JOURNAL. 

A  Monthly  Review  of  Veterinary  Science.  Established  1875.  Edited 
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the  Royal  Veterinary  College,  London.  Price  Is.  Annual  Subscription, 
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VETERINARY  NEWS. 

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